igm  (Thermo Fisher)


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  • 85
    Name:
    Mouse IgM Purified
    Description:
    • Isotype Mouse IgM• Class Class I ASR • Statement ASR Analyte Specific Reagent Analytical and performance characteristics are not established • Buffer PBS• Preservative 0 1 Sodium Azide
    Catalog Number:
    mgm00
    Price:
    None
    Applications:
    Cell Analysis|Flow Cytometry Antibodies & Secondary Detection|Flow Cytometry
    Category:
    Antibodies Secondary Detection Reagents
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    Structured Review

    Thermo Fisher igm
    Deposition of <t>IgM</t> in atherosclerotic lesions Confocal images of aortic root sections double-immunostained for <t>CD68</t> (green) and IgM (red) and counterstained with TOPRO nuclear dye (blue). (A) Low power view of aortic root lesion from LF-fed Ldlr −/− mouse showing IgM deposition in the acellular base of the lesion and its relationship to lesional macrophages. (B) Low power view of similar lesion from LF-fed sIgM.Ldlr −/− mouse showing complete absence of IgM.
    • Isotype Mouse IgM• Class Class I ASR • Statement ASR Analyte Specific Reagent Analytical and performance characteristics are not established • Buffer PBS• Preservative 0 1 Sodium Azide
    https://www.bioz.com/result/igm/product/Thermo Fisher
    Average 85 stars, based on 357 article reviews
    Price from $9.99 to $1999.99
    igm - by Bioz Stars, 2021-01
    85/100 stars

    Images

    1) Product Images from "Immunoglobulin M is Required for Protection Against Atherosclerosis in Low-Density Lipoprotein Receptor-Deficient Mice"

    Article Title: Immunoglobulin M is Required for Protection Against Atherosclerosis in Low-Density Lipoprotein Receptor-Deficient Mice

    Journal: Circulation

    doi: 10.1161/CIRCULATIONAHA.109.868158

    Deposition of IgM in atherosclerotic lesions Confocal images of aortic root sections double-immunostained for CD68 (green) and IgM (red) and counterstained with TOPRO nuclear dye (blue). (A) Low power view of aortic root lesion from LF-fed Ldlr −/− mouse showing IgM deposition in the acellular base of the lesion and its relationship to lesional macrophages. (B) Low power view of similar lesion from LF-fed sIgM.Ldlr −/− mouse showing complete absence of IgM.
    Figure Legend Snippet: Deposition of IgM in atherosclerotic lesions Confocal images of aortic root sections double-immunostained for CD68 (green) and IgM (red) and counterstained with TOPRO nuclear dye (blue). (A) Low power view of aortic root lesion from LF-fed Ldlr −/− mouse showing IgM deposition in the acellular base of the lesion and its relationship to lesional macrophages. (B) Low power view of similar lesion from LF-fed sIgM.Ldlr −/− mouse showing complete absence of IgM.

    Techniques Used:

    2) Product Images from "Peste des Petits Ruminants Virus-Like Particles Induce a Potent Humoral and Cellular Immune Response in Goats"

    Article Title: Peste des Petits Ruminants Virus-Like Particles Induce a Potent Humoral and Cellular Immune Response in Goats

    Journal: Viruses

    doi: 10.3390/v11100918

    Immunization induces a humoral response in goats. ( a ) Goats were boosted twice every three weeks via the subcutaneous route with VLPs (150 µg or 300 µg), PPRV Nigeria 75/1, PBS, or alum adjuvant after initial immunization. ( b ) Serum samples were collected 3, 6, 9, 12 and 15 weeks after the first vaccination and virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. ( c–d ) Total serum IgG (c) and IgM (d) responses were determined at 3 weeks. ( e–g ) Serum was collected from each goat three weeks after the third immunization for analysis by a PPRV F-, H-, and N-specific ELISA. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: *P
    Figure Legend Snippet: Immunization induces a humoral response in goats. ( a ) Goats were boosted twice every three weeks via the subcutaneous route with VLPs (150 µg or 300 µg), PPRV Nigeria 75/1, PBS, or alum adjuvant after initial immunization. ( b ) Serum samples were collected 3, 6, 9, 12 and 15 weeks after the first vaccination and virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. ( c–d ) Total serum IgG (c) and IgM (d) responses were determined at 3 weeks. ( e–g ) Serum was collected from each goat three weeks after the third immunization for analysis by a PPRV F-, H-, and N-specific ELISA. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: *P

    Techniques Used: Neutralization, Enzyme-linked Immunosorbent Assay

    Immunization induces a humoral immune response and Th1 class immunity in mice. ( a ) Mice were vaccinated three times via the subcutaneous route at 2-week intervals with VLPs, PPRV Nigeria 75/1 or PBS. Blood sampling and serum isolation were performed 2, 4, and 6 weeks after the first immunization. ( b ) Virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). ( c–i ) Serum was diluted by a factor of 5000, and total serum IgG and antibody subtypes were quantified by ELISA. The IgM (c) response were evaluated 1 week after the first vaccination, whereas IgA (d), IgG2a (e), IgG1 (f), IgG3 (g), IgG (h), and IgG2b (i) responses were determined 2 weeks after the second immunization. ( j ) The IgG1/IgG2a ratio was measured. Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: * P
    Figure Legend Snippet: Immunization induces a humoral immune response and Th1 class immunity in mice. ( a ) Mice were vaccinated three times via the subcutaneous route at 2-week intervals with VLPs, PPRV Nigeria 75/1 or PBS. Blood sampling and serum isolation were performed 2, 4, and 6 weeks after the first immunization. ( b ) Virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). ( c–i ) Serum was diluted by a factor of 5000, and total serum IgG and antibody subtypes were quantified by ELISA. The IgM (c) response were evaluated 1 week after the first vaccination, whereas IgA (d), IgG2a (e), IgG1 (f), IgG3 (g), IgG (h), and IgG2b (i) responses were determined 2 weeks after the second immunization. ( j ) The IgG1/IgG2a ratio was measured. Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: * P

    Techniques Used: Mouse Assay, Sampling, Isolation, Neutralization, Enzyme-linked Immunosorbent Assay

    3) Product Images from "Aryl Hydrocarbon Receptor Contributes to the Transcriptional Program of IL-10-Producing Regulatory B Cells"

    Article Title: Aryl Hydrocarbon Receptor Contributes to the Transcriptional Program of IL-10-Producing Regulatory B Cells

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2019.10.018

    CD19 + CD21 hi CD24 hi B Cells in Ahr fl/− Mb1 cre/+ Mice Are Less Able to Differentiate into Bregs AIA was induced in Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice. (A) Representative flow cytometry plots showing percentage and (B) bar charts showing the percentages and absolute numbers of CD19 + CD21 − CD24 hi (T1), CD19 + CD21 hi CD24 hi , and FO B cells in the spleens of Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice (n = 7). (C) CD19 + CD21 hi CD24 hi or FO B cells were sorted from Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice and stimulated with LPS+anti-IgM for 48 h. IL-10 production, as measured by ELISA (n = 4 per group). (D and E) Representative flow cytometry plots (D) showing percentage and (E) bar charts showing percentages and absolute numbers of CD19 + CD21 − CD24 hi , CD19 + CD21 hi CD24 hi and FO B cells in the MLNs of Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice (n = 7). (F) CD19 + B cells were sorted from Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice and stimulated with LPS+anti-IgM for 48 h. IL-10 production, as measured by ELISA (n = 3). All experiments were performed at day 7 post-IA injection. Data representative of at least two independent experiments with biological replicates. In (B), (C), (E), and (F), data are expressed as mean ± SEM. ∗ p
    Figure Legend Snippet: CD19 + CD21 hi CD24 hi B Cells in Ahr fl/− Mb1 cre/+ Mice Are Less Able to Differentiate into Bregs AIA was induced in Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice. (A) Representative flow cytometry plots showing percentage and (B) bar charts showing the percentages and absolute numbers of CD19 + CD21 − CD24 hi (T1), CD19 + CD21 hi CD24 hi , and FO B cells in the spleens of Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice (n = 7). (C) CD19 + CD21 hi CD24 hi or FO B cells were sorted from Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice and stimulated with LPS+anti-IgM for 48 h. IL-10 production, as measured by ELISA (n = 4 per group). (D and E) Representative flow cytometry plots (D) showing percentage and (E) bar charts showing percentages and absolute numbers of CD19 + CD21 − CD24 hi , CD19 + CD21 hi CD24 hi and FO B cells in the MLNs of Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice (n = 7). (F) CD19 + B cells were sorted from Mb1 cre/+ and Ahr fl/− Mb1 cre/+ mice and stimulated with LPS+anti-IgM for 48 h. IL-10 production, as measured by ELISA (n = 3). All experiments were performed at day 7 post-IA injection. Data representative of at least two independent experiments with biological replicates. In (B), (C), (E), and (F), data are expressed as mean ± SEM. ∗ p

    Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, IA, Injection

    4) Product Images from "Transgelin-2 in B-Cells Controls T-Cell Activation by Stabilizing T Cell - B Cell Conjugates"

    Article Title: Transgelin-2 in B-Cells Controls T-Cell Activation by Stabilizing T Cell - B Cell Conjugates

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0156429

    Transgelin-2-knockout mice exhibit normal B-cell development. (A) Expression of transgelin-2 in B-cells obtained from TAGLN2 +/+ (+/+) and TAGLN2 -/- (-/-) mice was determined by western blot analysis. M denotes molecular mass. (B) Single cell suspensions of bone marrow (BM) and spleen were tested for the presence of CD19 + and B220 + cells. The numbers in the dot plots indicate the percentage of CD19 + B220 + B-cells. The mean percentages of positive cells in BM and spleen are shown in the graph (right). (C) Single cell suspensions of BM from TAGLN2 +/+ and TAGLN2 -/- were assessed for the expression of CD43, CD21, CD25, IgM, and IgD by flow cytometry. The numbers in the dot plots indicate the percentage of B-cell subsets within the respective gates. The graph shows the average percentage of indicated B-cell development states in the BM cells (bottom). (D) Single cell suspensions of spleen from TAGLN2 +/+ and TAGLN2 -/- cells were pre-gated on B220, and expression of CD23, CD21, IgM, and IgD was assessed by flow cytometry. The numbers in the dot plots indicate the percentage of B-cell subsets within the respective gates. The graphs show the average percentage of the indicated B-cell subset in the B220 + B-cells gate (right). All dot plots are representative of two experiments with five mice, and the bar graphs are shown as means ± SD of two experiments with five mice each.
    Figure Legend Snippet: Transgelin-2-knockout mice exhibit normal B-cell development. (A) Expression of transgelin-2 in B-cells obtained from TAGLN2 +/+ (+/+) and TAGLN2 -/- (-/-) mice was determined by western blot analysis. M denotes molecular mass. (B) Single cell suspensions of bone marrow (BM) and spleen were tested for the presence of CD19 + and B220 + cells. The numbers in the dot plots indicate the percentage of CD19 + B220 + B-cells. The mean percentages of positive cells in BM and spleen are shown in the graph (right). (C) Single cell suspensions of BM from TAGLN2 +/+ and TAGLN2 -/- were assessed for the expression of CD43, CD21, CD25, IgM, and IgD by flow cytometry. The numbers in the dot plots indicate the percentage of B-cell subsets within the respective gates. The graph shows the average percentage of indicated B-cell development states in the BM cells (bottom). (D) Single cell suspensions of spleen from TAGLN2 +/+ and TAGLN2 -/- cells were pre-gated on B220, and expression of CD23, CD21, IgM, and IgD was assessed by flow cytometry. The numbers in the dot plots indicate the percentage of B-cell subsets within the respective gates. The graphs show the average percentage of the indicated B-cell subset in the B220 + B-cells gate (right). All dot plots are representative of two experiments with five mice, and the bar graphs are shown as means ± SD of two experiments with five mice each.

    Techniques Used: Knock-Out, Mouse Assay, Expressing, Western Blot, Flow Cytometry, Cytometry

    Transgelin-2 knockout had little effect on B-cell function. (A) Splenic B-cells obtained from TAGLN2 +/+ and TAGLN2 -/- mice were activated with anti-mouse IgM (10 μg/ml) for 12 h, and then CD69, MHCII, CD80, and CD86 expression were determined by flow cytometry. FACS histograms are representatives of three independent experiments, n = 3–6 mice per group. The bar graphs denote average mean fluorescence intensities (MFI) of each proteins ± SD. (B) B-cells were activated by PMA (200 nM) and ionomycin (1 μM), LPS (10 μg/ml), or IL-4 (1 μg/ml) and anti-CD40 (2 μg/ml). After 12 h, the CD69 expression level was assessed by flow cytometry (black line). FACS histograms are representatives of three independent experiments, n = 5 mice per group. The bar graph denotes average mean fluorescence intensities (MFI) ± SD. Gray shading shows isotype control. Veh, vehicle.
    Figure Legend Snippet: Transgelin-2 knockout had little effect on B-cell function. (A) Splenic B-cells obtained from TAGLN2 +/+ and TAGLN2 -/- mice were activated with anti-mouse IgM (10 μg/ml) for 12 h, and then CD69, MHCII, CD80, and CD86 expression were determined by flow cytometry. FACS histograms are representatives of three independent experiments, n = 3–6 mice per group. The bar graphs denote average mean fluorescence intensities (MFI) of each proteins ± SD. (B) B-cells were activated by PMA (200 nM) and ionomycin (1 μM), LPS (10 μg/ml), or IL-4 (1 μg/ml) and anti-CD40 (2 μg/ml). After 12 h, the CD69 expression level was assessed by flow cytometry (black line). FACS histograms are representatives of three independent experiments, n = 5 mice per group. The bar graph denotes average mean fluorescence intensities (MFI) ± SD. Gray shading shows isotype control. Veh, vehicle.

    Techniques Used: Knock-Out, Cell Function Assay, Mouse Assay, Expressing, Flow Cytometry, Cytometry, FACS, Fluorescence

    5) Product Images from "Assessment of enhanced influenza vaccination finds that FluAd conveys an advantage in mice and older adults"

    Article Title: Assessment of enhanced influenza vaccination finds that FluAd conveys an advantage in mice and older adults

    Journal: Clinical & Translational Immunology

    doi: 10.1002/cti2.1107

    Elevated CD4 + Tfh and GC B cells by increased expression of SHM and CSR genes lead to early IgG class switch for A‐eIIV. Vaccine responses were assessed by FACS (a) for CD4 + Tfh cells (b) and GC B cells (c) at days 7 and 21 post‐vaccination in the iLN. Expression of AID, Bcl6, IRF4, Pax5 and c‐Myc in B cells from iLN (relative expression to GAPDH expression, expressed in 2 Δ C t ) at day 7 post‐vaccination (d) . Quantity (f) and proportion (g) of H3N2‐1968 HA‐specific IgM and IgG at days 7 and 21 post‐vaccination, measured by ELISA AUC endpoint titres (Supplementary figure 2 ). Data represent the mean, SEM and individual responses, n = 4 or 5 mice per group. *shows statistical significance by one‐way ANOVA (mixed model) for eIIV versus S‐IIV, * P
    Figure Legend Snippet: Elevated CD4 + Tfh and GC B cells by increased expression of SHM and CSR genes lead to early IgG class switch for A‐eIIV. Vaccine responses were assessed by FACS (a) for CD4 + Tfh cells (b) and GC B cells (c) at days 7 and 21 post‐vaccination in the iLN. Expression of AID, Bcl6, IRF4, Pax5 and c‐Myc in B cells from iLN (relative expression to GAPDH expression, expressed in 2 Δ C t ) at day 7 post‐vaccination (d) . Quantity (f) and proportion (g) of H3N2‐1968 HA‐specific IgM and IgG at days 7 and 21 post‐vaccination, measured by ELISA AUC endpoint titres (Supplementary figure 2 ). Data represent the mean, SEM and individual responses, n = 4 or 5 mice per group. *shows statistical significance by one‐way ANOVA (mixed model) for eIIV versus S‐IIV, * P

    Techniques Used: Expressing, FACS, Enzyme-linked Immunosorbent Assay, Mouse Assay

    Memory B‐cell responses and long‐term antibody responses are increased in A‐eIIV‐vaccinated mice. BALB/c mice were vaccinated twice i.m. with either PBS, S‐IIV, A‐eIIV, H‐eIIV or R‐eIIV vaccines 3 weeks apart. iLN was collected at days 7 and 21 post‐vaccination to quantify by FACS (a) , IgG + (b) and IgM + (c) B memory cells. Longitudinal serum samples were serially sampled from individual mice at 2, 4 and 6 months post‐vaccination for H3N2‐2013 (d) , H3N2‐1968 (e) and H3‐stem (f) IgG by ELISA for avidity. Data represent the mean, SEM and individual responses, n = 4–8 mice per group. For (b, c) , *shows statistical significance for eIIV versus S‐IIV by one‐way ANOVA (mixed model) and Tukey's multiple comparison test. For d–f , # shows statistical significance for low‐avidity antibodies and *for high‐avidity antibodies for eIIV versus S‐IIV by one‐way ANOVA (mixed model) and Tukey's multiple comparison test. # or * P
    Figure Legend Snippet: Memory B‐cell responses and long‐term antibody responses are increased in A‐eIIV‐vaccinated mice. BALB/c mice were vaccinated twice i.m. with either PBS, S‐IIV, A‐eIIV, H‐eIIV or R‐eIIV vaccines 3 weeks apart. iLN was collected at days 7 and 21 post‐vaccination to quantify by FACS (a) , IgG + (b) and IgM + (c) B memory cells. Longitudinal serum samples were serially sampled from individual mice at 2, 4 and 6 months post‐vaccination for H3N2‐2013 (d) , H3N2‐1968 (e) and H3‐stem (f) IgG by ELISA for avidity. Data represent the mean, SEM and individual responses, n = 4–8 mice per group. For (b, c) , *shows statistical significance for eIIV versus S‐IIV by one‐way ANOVA (mixed model) and Tukey's multiple comparison test. For d–f , # shows statistical significance for low‐avidity antibodies and *for high‐avidity antibodies for eIIV versus S‐IIV by one‐way ANOVA (mixed model) and Tukey's multiple comparison test. # or * P

    Techniques Used: Mouse Assay, FACS, Enzyme-linked Immunosorbent Assay

    6) Product Images from "Heparosan-coated liposomes for drug delivery"

    Article Title: Heparosan-coated liposomes for drug delivery

    Journal: Glycobiology

    doi: 10.1093/glycob/cwx070

    Immunological challenge in rats with HEP-DiPalm and ELISA assessment. A set of three rats ( #1, 2, 3 ) then tested for the induction of an IgM or IgG response by ELISA (representative set of assays with averaged triplicate wells with standard deviation is presented). Overall, the signal from sera of rats before and after injection with HEP-DiPalm micelles (prelipid, white ; postlipid, black ) tested in the control wells coated with bovine serum albumin ( BSA ) was equivalent to the wells coated with HEP-BSA ( HEP ) or HEP-DiPalm ( DiPalm ) indicating that HEP is not significantly immunogenic, therefore, should be useful in multidose or long-term therapeutics.
    Figure Legend Snippet: Immunological challenge in rats with HEP-DiPalm and ELISA assessment. A set of three rats ( #1, 2, 3 ) then tested for the induction of an IgM or IgG response by ELISA (representative set of assays with averaged triplicate wells with standard deviation is presented). Overall, the signal from sera of rats before and after injection with HEP-DiPalm micelles (prelipid, white ; postlipid, black ) tested in the control wells coated with bovine serum albumin ( BSA ) was equivalent to the wells coated with HEP-BSA ( HEP ) or HEP-DiPalm ( DiPalm ) indicating that HEP is not significantly immunogenic, therefore, should be useful in multidose or long-term therapeutics.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Standard Deviation, Injection

    Representation of HEP-liposome assembly. Models of HEP-coated liposome assemblies with 13.3-kDa ( top ) and 7-kDa ( bottom ) HEP-lipid, based on typical molecular configurations from equilibrated coarse-grained simulations. The liposome and sugars are represented graphically by equivalent spheres (roughly to scale), and part of the liposome has been cut-away for viewing purposes. Potential interaction proteins are depicted (human serum albumin, HSA ; immunoglobulins IgG or IgM ; middle ), drawn to exact scale using coordinates from the Protein Data Bank. Less surface area is available for direct protein binding to liposomes coated with 0.5 mol% 13.3-kDa HEP-lipid compared to 0.5 mol% 7-kDa HEP-lipid, suggesting increased liposomal protection from clearance by the immune system. The image was rendered using POV-Ray software.
    Figure Legend Snippet: Representation of HEP-liposome assembly. Models of HEP-coated liposome assemblies with 13.3-kDa ( top ) and 7-kDa ( bottom ) HEP-lipid, based on typical molecular configurations from equilibrated coarse-grained simulations. The liposome and sugars are represented graphically by equivalent spheres (roughly to scale), and part of the liposome has been cut-away for viewing purposes. Potential interaction proteins are depicted (human serum albumin, HSA ; immunoglobulins IgG or IgM ; middle ), drawn to exact scale using coordinates from the Protein Data Bank. Less surface area is available for direct protein binding to liposomes coated with 0.5 mol% 13.3-kDa HEP-lipid compared to 0.5 mol% 7-kDa HEP-lipid, suggesting increased liposomal protection from clearance by the immune system. The image was rendered using POV-Ray software.

    Techniques Used: Protein Binding, Software

    7) Product Images from "Complete B Cell Deficiency Reduces Allograft Inflammation and Intragraft Macrophages a Rat Kidney Transplant Model"

    Article Title: Complete B Cell Deficiency Reduces Allograft Inflammation and Intragraft Macrophages a Rat Kidney Transplant Model

    Journal: Transplantation

    doi: 10.1097/TP.0000000000002010

    Immunophenotyping of B cell deficient rats demonstrated a lack of B cell populations and a lack of antibody production A and B, Flow cytometry demonstrated absence of B cells in lymphoid tissues in B −/− rats. Heterozygous (B +/− ) and wild type (WT) rats demonstrated similar B cell numbers. Systemic levels of IgM (C) and IgG (D) were undetectable in B −/− rats as measured by ELISA. WT and heterozygous rats had similar levels of circulating IgM and IgG. Similarly, splenocytes did not produce IgM or IgG as determined by ELISPOT analysis (E and F, respectively). * P
    Figure Legend Snippet: Immunophenotyping of B cell deficient rats demonstrated a lack of B cell populations and a lack of antibody production A and B, Flow cytometry demonstrated absence of B cells in lymphoid tissues in B −/− rats. Heterozygous (B +/− ) and wild type (WT) rats demonstrated similar B cell numbers. Systemic levels of IgM (C) and IgG (D) were undetectable in B −/− rats as measured by ELISA. WT and heterozygous rats had similar levels of circulating IgM and IgG. Similarly, splenocytes did not produce IgM or IgG as determined by ELISPOT analysis (E and F, respectively). * P

    Techniques Used: Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

    8) Product Images from "Spontaneous Production of Immunoglobulin M in Human Epithelial Cancer Cells"

    Article Title: Spontaneous Production of Immunoglobulin M in Human Epithelial Cancer Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0051423

    TLR9 agonists stimulated human epithelial cancer cells to secrete IgM. A, Ig µ mRNA level was analyzed by semiquantitative RT-PCR after stimulation by CpG 2006, and the two non-CpG ODN controls, CpG 2078 and GpC. CpG-N ODN208, as negative control; GAPDH, internal control. B, flow cytometry analysis showed that cytoplasmic IgM was decreased after stimulation with CpG 2006, CpG 2078, and GpC. CpG-N ODN208, as negative control. ** P
    Figure Legend Snippet: TLR9 agonists stimulated human epithelial cancer cells to secrete IgM. A, Ig µ mRNA level was analyzed by semiquantitative RT-PCR after stimulation by CpG 2006, and the two non-CpG ODN controls, CpG 2078 and GpC. CpG-N ODN208, as negative control; GAPDH, internal control. B, flow cytometry analysis showed that cytoplasmic IgM was decreased after stimulation with CpG 2006, CpG 2078, and GpC. CpG-N ODN208, as negative control. ** P

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Gel Permeation Chromatography, Negative Control, Flow Cytometry, Cytometry

    Chloroquine abolished effects for TLR9 agonists on IgM secretion in HeLa MR cells. A, semi-quantative RT-PCR showed increased Ig µ expression after stimulation by CpG 2006, CpG 2078 and GpC (upper panel). The effects of TLR9 agonists were abolished by chloroquine (lower panel). GAPDH, internal control. B, flow cytometry analysis showed that the cytoplasmic IgM level was decreased after stimulation with CpG 2006, CpG 2078 and GpC, and that this was abolished by chloroquine. * P
    Figure Legend Snippet: Chloroquine abolished effects for TLR9 agonists on IgM secretion in HeLa MR cells. A, semi-quantative RT-PCR showed increased Ig µ expression after stimulation by CpG 2006, CpG 2078 and GpC (upper panel). The effects of TLR9 agonists were abolished by chloroquine (lower panel). GAPDH, internal control. B, flow cytometry analysis showed that the cytoplasmic IgM level was decreased after stimulation with CpG 2006, CpG 2078 and GpC, and that this was abolished by chloroquine. * P

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Gel Permeation Chromatography, Flow Cytometry, Cytometry

    Knockdown of MyD88 by siRNA inhibited TLR9 agonist-induced IgM secretion in HeLa MR cells. A, effects of three synthesized siRNAs for knocking down MyD88 expression were analyzed after transfection from 24 h to 96 h by semiquantitative RT-PCR. Results of the effective siRNAs 1 and 3 are shown. B, semi-quantative RT-PCR showed increased Ig µ expression after stimulation by CpG 2006, CpG 2078 and GpC (upper panel). This was abolished by MyD88 knockdown with siRNA1 and siRNA3 (lower panel). GAPDH, internal control. C, flow cytometry analysis showed that the cytoplasmic IgM level was decreased after stimulation with CpG 2006, CpG 2078 and GpC, and that this was abolished by MyD88 knockdown with siRNA1 and siRNA3. ** P
    Figure Legend Snippet: Knockdown of MyD88 by siRNA inhibited TLR9 agonist-induced IgM secretion in HeLa MR cells. A, effects of three synthesized siRNAs for knocking down MyD88 expression were analyzed after transfection from 24 h to 96 h by semiquantitative RT-PCR. Results of the effective siRNAs 1 and 3 are shown. B, semi-quantative RT-PCR showed increased Ig µ expression after stimulation by CpG 2006, CpG 2078 and GpC (upper panel). This was abolished by MyD88 knockdown with siRNA1 and siRNA3 (lower panel). GAPDH, internal control. C, flow cytometry analysis showed that the cytoplasmic IgM level was decreased after stimulation with CpG 2006, CpG 2078 and GpC, and that this was abolished by MyD88 knockdown with siRNA1 and siRNA3. ** P

    Techniques Used: Synthesized, Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Gel Permeation Chromatography, Flow Cytometry, Cytometry

    9) Product Images from "Immune Tolerance to Apoptotic Self Is Mediated Primarily by Regulatory B1a Cells"

    Article Title: Immune Tolerance to Apoptotic Self Is Mediated Primarily by Regulatory B1a Cells

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.01952

    IL-10 +ve apoptotic cell (AC) responsive B cells express CD43 + CD24 hi CD21 hi CD23 lo IgD lo IgM hi . (A) Splenic CD19 +ve cells were FACS sorted (i) into CD24 lo CD21 lo (A1; 69.8% of all B cells) and CD24 hi CD21 hi (A2; 9.8% of all B cells). A2 was further sorted into CD23 lo (A3 50.8% of A2) and CD23 hi A in Supplementary Material for cell purity. (B) Splenic CD19 +ve cells were FACS sorted (i) into IgD hi IgM lo (C1 52.5% of all B cells) and IgD lo IgM hi (C2 13.1% of all B cells). C2 was further sorted into CD21 lo (C3 49.7% of C2) and CD21 hi B in Supplementary Material for cell purity. (C) Apoptotic cells were injected IV on D0, 2, and 5 into IL-10-GFP expressing mice. Splenic CD19 +ve cells were isolated on D7. (i) IL-10 +ve B cells. (ii) Surface markers of IL-10-GFP +ve B cells (in red) and IL-10-GFP −ve B cells (in black) compared to isotype control (in shaded gray). (iii) The fold difference in mean florescence intensity (MFI) (IL-10-GFP +ve versus IL-10-GFP −ve C in Supplementary Material for sorting strategy and cell purity. (D) CD19 +ve B cells were sorted into MZB (CD19 +ve CD23 lo CD21 hi CD43 −ve ), B1a (CD19 +ve CD23 lo CD21 hi CD43 +ve ) B cells, and follicular B (FOB) cells (CD19 +ve CD23 +ve CD21 lo CD43 −ve ). Cells were cultured with 2 μg/ml OVA peptide, DO11.10 CD4 + T cells ± apoptotic cells. After 72 h cells, the total IL-10 secreted by stimulated B and T cells was measured by ELISA. (E) Apoptotic cells were injected IV on D0, 2, and 5 into IL-10-GFP expressing mice. Peritoneal cavity CD19 +ve cells were isolated on D7. (i) IL-10 +ve B cells. (ii) Surface markers of IL-10-GFP +ve B cells (in red) and IL-10-GFP −ve B cells (in black) compared to isotype control (in shaded gray). (iii) The fold difference in MFI (IL-10-GFP +ve versus IL-10-GFP −ve B cells), is shown for a range of other surface markers. (F) CD43 +ve splenic and peritoneal B1 B cells express higher levels of transcription factor Bhlhe41, when compared to splenic FOB and marginal zone B cells or peritoneal CD43 −ve B cells. Data were normalized using 18S then expressed relative to FOB. (G) B cell subsets were sorted into splenic FOB (CD23 hi CD21 lo CD43 −ve ), peritoneal cavity B1a (CD19 hi CD43 +ve CD5 +ve ) and B1b (CD19 hi CD43 +ve CD5 −ve ) cells. Sorted cells were cultured in the presence of apoptotic cells and R848 at varying concentrations. Cells were stained after 72 h for intracellular CD19 +ve IL-10. All experiments are representative of three individual experiments [except (C,D) , where FACs data are representative of 13 individual experiments. Activation marker changes are graphed from pooled data of min six individual mice]. Statistical differences were determine by unpaired Student’s t -test * P
    Figure Legend Snippet: IL-10 +ve apoptotic cell (AC) responsive B cells express CD43 + CD24 hi CD21 hi CD23 lo IgD lo IgM hi . (A) Splenic CD19 +ve cells were FACS sorted (i) into CD24 lo CD21 lo (A1; 69.8% of all B cells) and CD24 hi CD21 hi (A2; 9.8% of all B cells). A2 was further sorted into CD23 lo (A3 50.8% of A2) and CD23 hi A in Supplementary Material for cell purity. (B) Splenic CD19 +ve cells were FACS sorted (i) into IgD hi IgM lo (C1 52.5% of all B cells) and IgD lo IgM hi (C2 13.1% of all B cells). C2 was further sorted into CD21 lo (C3 49.7% of C2) and CD21 hi B in Supplementary Material for cell purity. (C) Apoptotic cells were injected IV on D0, 2, and 5 into IL-10-GFP expressing mice. Splenic CD19 +ve cells were isolated on D7. (i) IL-10 +ve B cells. (ii) Surface markers of IL-10-GFP +ve B cells (in red) and IL-10-GFP −ve B cells (in black) compared to isotype control (in shaded gray). (iii) The fold difference in mean florescence intensity (MFI) (IL-10-GFP +ve versus IL-10-GFP −ve C in Supplementary Material for sorting strategy and cell purity. (D) CD19 +ve B cells were sorted into MZB (CD19 +ve CD23 lo CD21 hi CD43 −ve ), B1a (CD19 +ve CD23 lo CD21 hi CD43 +ve ) B cells, and follicular B (FOB) cells (CD19 +ve CD23 +ve CD21 lo CD43 −ve ). Cells were cultured with 2 μg/ml OVA peptide, DO11.10 CD4 + T cells ± apoptotic cells. After 72 h cells, the total IL-10 secreted by stimulated B and T cells was measured by ELISA. (E) Apoptotic cells were injected IV on D0, 2, and 5 into IL-10-GFP expressing mice. Peritoneal cavity CD19 +ve cells were isolated on D7. (i) IL-10 +ve B cells. (ii) Surface markers of IL-10-GFP +ve B cells (in red) and IL-10-GFP −ve B cells (in black) compared to isotype control (in shaded gray). (iii) The fold difference in MFI (IL-10-GFP +ve versus IL-10-GFP −ve B cells), is shown for a range of other surface markers. (F) CD43 +ve splenic and peritoneal B1 B cells express higher levels of transcription factor Bhlhe41, when compared to splenic FOB and marginal zone B cells or peritoneal CD43 −ve B cells. Data were normalized using 18S then expressed relative to FOB. (G) B cell subsets were sorted into splenic FOB (CD23 hi CD21 lo CD43 −ve ), peritoneal cavity B1a (CD19 hi CD43 +ve CD5 +ve ) and B1b (CD19 hi CD43 +ve CD5 −ve ) cells. Sorted cells were cultured in the presence of apoptotic cells and R848 at varying concentrations. Cells were stained after 72 h for intracellular CD19 +ve IL-10. All experiments are representative of three individual experiments [except (C,D) , where FACs data are representative of 13 individual experiments. Activation marker changes are graphed from pooled data of min six individual mice]. Statistical differences were determine by unpaired Student’s t -test * P

    Techniques Used: FACS, Injection, Expressing, Mouse Assay, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay, Staining, Activation Assay, Marker

    Regulatory B cells that recognize apoptotic cells, produce both IL-10 and IgM autoantibodies. (A) Ai in Supplementary Material. (ii) Cells were also visualized by confocal microscopy to show B/AC interaction. IgM +ve Aii in Supplementary Material. Data representative of two separate experiments. (B) ELISA of serum IgM from mice given AC infusions 7 days earlier. Naive untreated mice are shown in the patterned bars and AC-treated mice in the solid bars. Data pooled of five individual mice and from two experiments. (C) IL-10 +ve and IL-10 −ve CD19 +ve B cells were isolated from spleens on D7 post-AC injection and cultured with MegaAPRIL, IL-4, and CpG. (i) Supernatants were tested for IgM on Day 10. Data are representative of n = 3. (ii) ELISA of secreted IgM from (C) (i). IgM from IL-10 −ve cells is shown with patterned bars and IL10 +ve cells with solid bars. Data representative of n = 3 and pooled from two individual experiments. (D) CD19 +ve IL-10 +ve D in Supplementary Material). Positive binding clones were then checked for IgM to specified autoantigens using an equal concentration of IgM per clone tested. Binding from IL-10 −ve B cell clones were used as a negative control (shown with dotted line on each graph). No IgG was detected (data now shown). Positive control (red line) serum from MRL-lpr/lpr lupus mouse was included. Data are pooled from three individual experiments.
    Figure Legend Snippet: Regulatory B cells that recognize apoptotic cells, produce both IL-10 and IgM autoantibodies. (A) Ai in Supplementary Material. (ii) Cells were also visualized by confocal microscopy to show B/AC interaction. IgM +ve Aii in Supplementary Material. Data representative of two separate experiments. (B) ELISA of serum IgM from mice given AC infusions 7 days earlier. Naive untreated mice are shown in the patterned bars and AC-treated mice in the solid bars. Data pooled of five individual mice and from two experiments. (C) IL-10 +ve and IL-10 −ve CD19 +ve B cells were isolated from spleens on D7 post-AC injection and cultured with MegaAPRIL, IL-4, and CpG. (i) Supernatants were tested for IgM on Day 10. Data are representative of n = 3. (ii) ELISA of secreted IgM from (C) (i). IgM from IL-10 −ve cells is shown with patterned bars and IL10 +ve cells with solid bars. Data representative of n = 3 and pooled from two individual experiments. (D) CD19 +ve IL-10 +ve D in Supplementary Material). Positive binding clones were then checked for IgM to specified autoantigens using an equal concentration of IgM per clone tested. Binding from IL-10 −ve B cell clones were used as a negative control (shown with dotted line on each graph). No IgG was detected (data now shown). Positive control (red line) serum from MRL-lpr/lpr lupus mouse was included. Data are pooled from three individual experiments.

    Techniques Used: Confocal Microscopy, Enzyme-linked Immunosorbent Assay, Mouse Assay, Isolation, Injection, Cell Culture, Binding Assay, Clone Assay, Concentration Assay, Negative Control, Positive Control

    10) Product Images from "TGF-β-Induced CD8+CD103+ Regulatory T Cells Show Potent Therapeutic Effect on Chronic Graft-versus-Host Disease Lupus by Suppressing B Cells"

    Article Title: TGF-β-Induced CD8+CD103+ Regulatory T Cells Show Potent Therapeutic Effect on Chronic Graft-versus-Host Disease Lupus by Suppressing B Cells

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.00035

    CD8 + CD103 + iTregs directly suppress B cell proliferation and the ability to secrete antibody ex vivo . (A) Fresh B cells labeled with CFSE were cultured in 96-well plates with CD8 + CD103 − med, CD8 + CD103 + iTreg, CD4 + iTreg, or nTreg cells (the ratio of T cells: B cells was 1:2) in the presence of lipopolysaccharide. B cell proliferation was determined by the CFSE dilution rates after 3 days of culture. Typical FACS histogram and summary data were shown. (B) The supernatants were collected from T cells and B cells co-culture systems after 3-day culture, and the IgG and IgM secretion was detected by ELISA. The data indicate the mean ± SEM of three independent experiments (NS means no significance, * P
    Figure Legend Snippet: CD8 + CD103 + iTregs directly suppress B cell proliferation and the ability to secrete antibody ex vivo . (A) Fresh B cells labeled with CFSE were cultured in 96-well plates with CD8 + CD103 − med, CD8 + CD103 + iTreg, CD4 + iTreg, or nTreg cells (the ratio of T cells: B cells was 1:2) in the presence of lipopolysaccharide. B cell proliferation was determined by the CFSE dilution rates after 3 days of culture. Typical FACS histogram and summary data were shown. (B) The supernatants were collected from T cells and B cells co-culture systems after 3-day culture, and the IgG and IgM secretion was detected by ELISA. The data indicate the mean ± SEM of three independent experiments (NS means no significance, * P

    Techniques Used: Ex Vivo, Labeling, Cell Culture, FACS, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

    11) Product Images from "Small ?2-glycoprotein I peptides protect from intestinal ischemia reperfusion injury"

    Article Title: Small ?2-glycoprotein I peptides protect from intestinal ischemia reperfusion injury

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1200290

    Peptide Retro D-p9 attenuate IgM and C3 deposition in response to I/R Prior to Sham or I/R treatment C57Bl/6 mice were treated with or without injection of 40 μM β2-GPI peptides, p9, D-p9 or Retro D-p9. Representative intestinal sections stained for IgM (A) or C3 (B) deposition. Additional sections were stained for myeloperoxidase (C). Fluorescent intensity was quantitated using Image J with threshold set on isotype control tissues. Each bar is the average ± SEM with 5–8 animals per treatment and 4–6 photographs per animal. Magnification for all photomicrographs and measurements were obtained at 200x. Microphotographs are representative of 3–4 animals stained in at least 3 independent experiments. * = p ≤ 0.05 compared to Sham + peptide, Φ = p ≤ 0.05 compared to I/R treatment animals not receiving peptides.
    Figure Legend Snippet: Peptide Retro D-p9 attenuate IgM and C3 deposition in response to I/R Prior to Sham or I/R treatment C57Bl/6 mice were treated with or without injection of 40 μM β2-GPI peptides, p9, D-p9 or Retro D-p9. Representative intestinal sections stained for IgM (A) or C3 (B) deposition. Additional sections were stained for myeloperoxidase (C). Fluorescent intensity was quantitated using Image J with threshold set on isotype control tissues. Each bar is the average ± SEM with 5–8 animals per treatment and 4–6 photographs per animal. Magnification for all photomicrographs and measurements were obtained at 200x. Microphotographs are representative of 3–4 animals stained in at least 3 independent experiments. * = p ≤ 0.05 compared to Sham + peptide, Φ = p ≤ 0.05 compared to I/R treatment animals not receiving peptides.

    Techniques Used: Mouse Assay, Injection, Staining

    12) Product Images from "A novel IgM–H-Ficolin complement pathway to attack allogenic cancer cells in vitro"

    Article Title: A novel IgM–H-Ficolin complement pathway to attack allogenic cancer cells in vitro

    Journal: Scientific Reports

    doi: 10.1038/srep07824

    IgM but not IgG was involved in H-ficolin–mediated complement activation pathway. (a) The purified IgG/albumin (P-IgG/A) and the IgG/albumin-depleted PHS or NHS (IgG/A-depleted PHS or NHS) were stained by C. Blue or analyzed by Western blotting. (b) The cell lysing and binding activity of P-IgG/A and IgG/A-depleted PHS were determined. (c) The IgM-depleted PHS or NHS (IgM-depleted PHS or NHS) and the purified IgM from PHS (P-IgM) or NHS (N-IgM) were analyzed. (d) The cell lysing and binding activity of IgM-depleted PHS and P-IgM. (e) Schematic illustration of the IgM–H-ficolin signal pathway in complement activation. (NS indicates p > 0.05, * p
    Figure Legend Snippet: IgM but not IgG was involved in H-ficolin–mediated complement activation pathway. (a) The purified IgG/albumin (P-IgG/A) and the IgG/albumin-depleted PHS or NHS (IgG/A-depleted PHS or NHS) were stained by C. Blue or analyzed by Western blotting. (b) The cell lysing and binding activity of P-IgG/A and IgG/A-depleted PHS were determined. (c) The IgM-depleted PHS or NHS (IgM-depleted PHS or NHS) and the purified IgM from PHS (P-IgM) or NHS (N-IgM) were analyzed. (d) The cell lysing and binding activity of IgM-depleted PHS and P-IgM. (e) Schematic illustration of the IgM–H-ficolin signal pathway in complement activation. (NS indicates p > 0.05, * p

    Techniques Used: Activation Assay, Purification, Staining, Western Blot, Binding Assay, Activity Assay

    13) Product Images from "Combined proinflammatory cytokine and cognate activation of invariant natural killer T cells enhances anti-DNA antibody responses"

    Article Title: Combined proinflammatory cytokine and cognate activation of invariant natural killer T cells enhances anti-DNA antibody responses

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1920463117

    αGalCer-mediated i NKT cell activation enhances self-reactive innate B cell responses in chronic inflammation. ( A ) Experimental scheme. Mice were immunized i.p. with 5 μg α-galactosylceramide (αGC) on day 1 and i.p. 2 μg IL-18 for days 1 to 10. To follow NP-specific B cell responses, 0.5 μg of NP-αGC was injected intravenously. ( B ) ELISA was used to measure total Ig concentrations from serum at the indicated time points (the statistical difference between IL-18 and IL-18 + αGalCer is shown). ( C ) OD of anti-DNA IgE, IgM, and IgG on day 12. ( D ) Anti-DNA IgG1, IgG2b, and IgG3 and the ratio of IgG2b vs. IgG1 on day 12. Two-way ( B ) or one-way ANOVA ( C and D ) was employed to assess statistical differences. * P
    Figure Legend Snippet: αGalCer-mediated i NKT cell activation enhances self-reactive innate B cell responses in chronic inflammation. ( A ) Experimental scheme. Mice were immunized i.p. with 5 μg α-galactosylceramide (αGC) on day 1 and i.p. 2 μg IL-18 for days 1 to 10. To follow NP-specific B cell responses, 0.5 μg of NP-αGC was injected intravenously. ( B ) ELISA was used to measure total Ig concentrations from serum at the indicated time points (the statistical difference between IL-18 and IL-18 + αGalCer is shown). ( C ) OD of anti-DNA IgE, IgM, and IgG on day 12. ( D ) Anti-DNA IgG1, IgG2b, and IgG3 and the ratio of IgG2b vs. IgG1 on day 12. Two-way ( B ) or one-way ANOVA ( C and D ) was employed to assess statistical differences. * P

    Techniques Used: Activation Assay, Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay

    Cognate activation of i NKT cells during chronic inflammation provides B cell help and IL-18 + αGalCer exacerbates collagen-induced arthritis. ( A and B ) Percent of ( A ) GFP+ plasma cells and ( B ) hCD2+ germinal center B cells from spleens of Aicda Cre GT Rosa Flox mice treated as indicated. Data are representative of three experiments ( n = 3 mice per group). ( C ) OD (450 nm) of NP-reactive IgM, IgG, IgG1, and IgG3 measured by ELISA. Sera were diluted as shown on the x axis. Data are representative of two experiments ( n = 4 or 5 mice per group). ( D ) Percent of intracellular, NP24-reactive IgG3+ cells among plasma cells (CD138+, B220−). ( E ) Representative flow cytometry plots for intracellular staining. ( F ) Representative immunofluorescence microscopy of the spleen (day 12) showing B220 (blue), hCD2 (green), and IgG3 (red) antibody-forming cells. (Scale bars, 100 μm.) Data are representative of two experiments ( n = 4 or 5 mice per group). One-way ( A , B , D ) or two-way ( C ) ANOVA, Student's t test ( H ). *P
    Figure Legend Snippet: Cognate activation of i NKT cells during chronic inflammation provides B cell help and IL-18 + αGalCer exacerbates collagen-induced arthritis. ( A and B ) Percent of ( A ) GFP+ plasma cells and ( B ) hCD2+ germinal center B cells from spleens of Aicda Cre GT Rosa Flox mice treated as indicated. Data are representative of three experiments ( n = 3 mice per group). ( C ) OD (450 nm) of NP-reactive IgM, IgG, IgG1, and IgG3 measured by ELISA. Sera were diluted as shown on the x axis. Data are representative of two experiments ( n = 4 or 5 mice per group). ( D ) Percent of intracellular, NP24-reactive IgG3+ cells among plasma cells (CD138+, B220−). ( E ) Representative flow cytometry plots for intracellular staining. ( F ) Representative immunofluorescence microscopy of the spleen (day 12) showing B220 (blue), hCD2 (green), and IgG3 (red) antibody-forming cells. (Scale bars, 100 μm.) Data are representative of two experiments ( n = 4 or 5 mice per group). One-way ( A , B , D ) or two-way ( C ) ANOVA, Student's t test ( H ). *P

    Techniques Used: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Staining, Immunofluorescence, Microscopy

    14) Product Images from "Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers"

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers

    Journal: Scientific Reports

    doi: 10.1038/srep30004

    IL-6 and LPS activate IgM secretion in naïve B cells. ( a ) ELISPOT analysis of IgM-secreting cells in splenocyte cultures treated with IL-6 (200 ng/ml), LPS (100 μg/ml) or non-stimulated. Splenocytes were cultured for 3 days in ELISPOT plates previously coated with anti-trout IgM mAb (2 μg/ml) in the presence or absence of the different stimuli. After incubation, cells were washed away and a biotinylated anti-trout IgM mAb (1 μg/ml) was used to detect numbers of spot forming cells. Duplicates from a representative experiment (left) and quantification of spot forming cells (right) from 5 independent experiments are shown (mean + standard deviation). ( b ) Spleen leukocytes were incubated with media containing IL-6, LPS or control media alone for 24 h at 20 °C. After that time, IgM + B cells were sorted using an anti-trout IgM mAb and RNA was extracted. Relative transcript expression of Blimp-1 and ACKR2 is shown (mean + standard deviation, n = 6). ( c ) Membrane IgM and total IgM expression of IgM + cells after incubation with IL-6, LPS or control media for 24, 48 and 72 h. Mean fluorescence intensity (MFI) + standard deviation is shown (n = 9). ( d ) Dot plots and histograms showing the Forward scatter (FSC) from IgM + B cells and BrdU + /IgM + B cells incubated in the presence or absence of LPS or IL-6, from one representative experiment. Graphs showing FSC MFI values from 8 independent experiments (mean + standard deviation) are included next to the histograms for stimulated cultures. * P
    Figure Legend Snippet: IL-6 and LPS activate IgM secretion in naïve B cells. ( a ) ELISPOT analysis of IgM-secreting cells in splenocyte cultures treated with IL-6 (200 ng/ml), LPS (100 μg/ml) or non-stimulated. Splenocytes were cultured for 3 days in ELISPOT plates previously coated with anti-trout IgM mAb (2 μg/ml) in the presence or absence of the different stimuli. After incubation, cells were washed away and a biotinylated anti-trout IgM mAb (1 μg/ml) was used to detect numbers of spot forming cells. Duplicates from a representative experiment (left) and quantification of spot forming cells (right) from 5 independent experiments are shown (mean + standard deviation). ( b ) Spleen leukocytes were incubated with media containing IL-6, LPS or control media alone for 24 h at 20 °C. After that time, IgM + B cells were sorted using an anti-trout IgM mAb and RNA was extracted. Relative transcript expression of Blimp-1 and ACKR2 is shown (mean + standard deviation, n = 6). ( c ) Membrane IgM and total IgM expression of IgM + cells after incubation with IL-6, LPS or control media for 24, 48 and 72 h. Mean fluorescence intensity (MFI) + standard deviation is shown (n = 9). ( d ) Dot plots and histograms showing the Forward scatter (FSC) from IgM + B cells and BrdU + /IgM + B cells incubated in the presence or absence of LPS or IL-6, from one representative experiment. Graphs showing FSC MFI values from 8 independent experiments (mean + standard deviation) are included next to the histograms for stimulated cultures. * P

    Techniques Used: Enzyme-linked Immunospot, Cell Culture, Incubation, Standard Deviation, Expressing, Fluorescence

    15) Product Images from "Peste des Petits Ruminants Virus-Like Particles Induce a Potent Humoral and Cellular Immune Response in Goats"

    Article Title: Peste des Petits Ruminants Virus-Like Particles Induce a Potent Humoral and Cellular Immune Response in Goats

    Journal: Viruses

    doi: 10.3390/v11100918

    Immunization induces a humoral response in goats. ( a ) Goats were boosted twice every three weeks via the subcutaneous route with VLPs (150 µg or 300 µg), PPRV Nigeria 75/1, PBS, or alum adjuvant after initial immunization. ( b ) Serum samples were collected 3, 6, 9, 12 and 15 weeks after the first vaccination and virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. ( c–d ) Total serum IgG (c) and IgM (d) responses were determined at 3 weeks. ( e–g ) Serum was collected from each goat three weeks after the third immunization for analysis by a PPRV F-, H-, and N-specific ELISA. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: *P
    Figure Legend Snippet: Immunization induces a humoral response in goats. ( a ) Goats were boosted twice every three weeks via the subcutaneous route with VLPs (150 µg or 300 µg), PPRV Nigeria 75/1, PBS, or alum adjuvant after initial immunization. ( b ) Serum samples were collected 3, 6, 9, 12 and 15 weeks after the first vaccination and virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. ( c–d ) Total serum IgG (c) and IgM (d) responses were determined at 3 weeks. ( e–g ) Serum was collected from each goat three weeks after the third immunization for analysis by a PPRV F-, H-, and N-specific ELISA. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: *P

    Techniques Used: Neutralization, Enzyme-linked Immunosorbent Assay

    Immunization induces a humoral immune response and Th1 class immunity in mice. ( a ) Mice were vaccinated three times via the subcutaneous route at 2-week intervals with VLPs, PPRV Nigeria 75/1 or PBS. Blood sampling and serum isolation were performed 2, 4, and 6 weeks after the first immunization. ( b ) Virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). ( c–i ) Serum was diluted by a factor of 5000, and total serum IgG and antibody subtypes were quantified by ELISA. The IgM (c) response were evaluated 1 week after the first vaccination, whereas IgA (d), IgG2a (e), IgG1 (f), IgG3 (g), IgG (h), and IgG2b (i) responses were determined 2 weeks after the second immunization. ( j ) The IgG1/IgG2a ratio was measured. Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: * P
    Figure Legend Snippet: Immunization induces a humoral immune response and Th1 class immunity in mice. ( a ) Mice were vaccinated three times via the subcutaneous route at 2-week intervals with VLPs, PPRV Nigeria 75/1 or PBS. Blood sampling and serum isolation were performed 2, 4, and 6 weeks after the first immunization. ( b ) Virus-neutralizing antibody (VNA) titers were measured by fluorescent antibody viral neutralization (FAVN). ( c–i ) Serum was diluted by a factor of 5000, and total serum IgG and antibody subtypes were quantified by ELISA. The IgM (c) response were evaluated 1 week after the first vaccination, whereas IgA (d), IgG2a (e), IgG1 (f), IgG3 (g), IgG (h), and IgG2b (i) responses were determined 2 weeks after the second immunization. ( j ) The IgG1/IgG2a ratio was measured. Dotted lines represent antibody titers greater than ten, indicating positive serum conversion. Significant differences were analyzed by one-way or two-way ANOVA and indicated as follows: * P

    Techniques Used: Mouse Assay, Sampling, Isolation, Neutralization, Enzyme-linked Immunosorbent Assay

    16) Product Images from "Tick-host conflict: immunoglobulin E antibodies to tick proteins in patients with anaphylaxis to tick bite"

    Article Title: Tick-host conflict: immunoglobulin E antibodies to tick proteins in patients with anaphylaxis to tick bite

    Journal: Oncotarget

    doi: 10.18632/oncotarget.15243

    Immunological response to tick proteins The IgE, IgM and IgG antibody levels were determined by ELISA in patients and control serum samples against A . α-Gal, B . R. bursa salivary gland proteins, C . H. marginatum salivary gland proteins, and D . R. microplus BME/CTVM23 tick cell proteins. Antibody levels were determined as OD at 450 nm and shown as average + SD of 4 technical replicates.
    Figure Legend Snippet: Immunological response to tick proteins The IgE, IgM and IgG antibody levels were determined by ELISA in patients and control serum samples against A . α-Gal, B . R. bursa salivary gland proteins, C . H. marginatum salivary gland proteins, and D . R. microplus BME/CTVM23 tick cell proteins. Antibody levels were determined as OD at 450 nm and shown as average + SD of 4 technical replicates.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Patient-specific antibody response to tick species responsible for the reported anaphylactic reaction to tick bite A . Correlation analysis between IgE, IgM and IgG antibody levels against R. bursa or H. marginatum tick proteins and α-Gal in patients 1 and 2 and healthy control individual. Antibody levels were determined as OD at 450 nm and shown as the average of 4 technical replicates. B . The IgE response to R. bursa and H. marginatum salivary gland and R. microplus BME/CTVM23 tick cell proteins was analyzed by 1-D Western blot using patient 1 and 2 sera. Abbreviation: MW, molecular weight protein marker.
    Figure Legend Snippet: Patient-specific antibody response to tick species responsible for the reported anaphylactic reaction to tick bite A . Correlation analysis between IgE, IgM and IgG antibody levels against R. bursa or H. marginatum tick proteins and α-Gal in patients 1 and 2 and healthy control individual. Antibody levels were determined as OD at 450 nm and shown as the average of 4 technical replicates. B . The IgE response to R. bursa and H. marginatum salivary gland and R. microplus BME/CTVM23 tick cell proteins was analyzed by 1-D Western blot using patient 1 and 2 sera. Abbreviation: MW, molecular weight protein marker.

    Techniques Used: Western Blot, Molecular Weight, Marker

    17) Product Images from "Heparanase-neutralizing antibodies attenuate lymphoma tumor growth and metastasis"

    Article Title: Heparanase-neutralizing antibodies attenuate lymphoma tumor growth and metastasis

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1519453113

    Heparanase neutralizing mAbs attenuate cell invasion and tumor metastasis. ( A ) Cell invasion. Cal-27 tongue carcinoma ( Left ) and U87 glioma cells ( Middle ) (2 × 10 5 ) were plated onto Matrigel-coated filters in the presence of mouse IgM/IgG ( Upper
    Figure Legend Snippet: Heparanase neutralizing mAbs attenuate cell invasion and tumor metastasis. ( A ) Cell invasion. Cal-27 tongue carcinoma ( Left ) and U87 glioma cells ( Middle ) (2 × 10 5 ) were plated onto Matrigel-coated filters in the presence of mouse IgM/IgG ( Upper

    Techniques Used:

    18) Product Images from "A Novel Targeted Screening Tool for Hypogammaglobulinemia: Measurement of Serum Immunoglobulin (IgG, IgM, IgA) Levels from Dried Blood Spots (Ig-DBS Assay)"

    Article Title: A Novel Targeted Screening Tool for Hypogammaglobulinemia: Measurement of Serum Immunoglobulin (IgG, IgM, IgA) Levels from Dried Blood Spots (Ig-DBS Assay)

    Journal: Journal of Clinical Immunology

    doi: 10.1007/s10875-015-0184-y

    Correlations between standard nephelometry assay and Ig-DBS for IgG ( a ), IgM ( b ), and IgA ( c ) ( scatterplot and regression line )
    Figure Legend Snippet: Correlations between standard nephelometry assay and Ig-DBS for IgG ( a ), IgM ( b ), and IgA ( c ) ( scatterplot and regression line )

    Techniques Used: Nephelometric Assay

    19) Product Images from "A Novel Targeted Screening Tool for Hypogammaglobulinemia: Measurement of Serum Immunoglobulin (IgG, IgM, IgA) Levels from Dried Blood Spots (Ig-DBS Assay)"

    Article Title: A Novel Targeted Screening Tool for Hypogammaglobulinemia: Measurement of Serum Immunoglobulin (IgG, IgM, IgA) Levels from Dried Blood Spots (Ig-DBS Assay)

    Journal: Journal of Clinical Immunology

    doi: 10.1007/s10875-015-0184-y

    Correlations between standard nephelometry assay and Ig-DBS for IgG ( a ), IgM ( b ), and IgA ( c ) ( scatterplot and regression line )
    Figure Legend Snippet: Correlations between standard nephelometry assay and Ig-DBS for IgG ( a ), IgM ( b ), and IgA ( c ) ( scatterplot and regression line )

    Techniques Used: Nephelometric Assay

    20) Product Images from "Plasmodium chabaudi infection induces AID expression in transitional and marginal zone B cells"

    Article Title: Plasmodium chabaudi infection induces AID expression in transitional and marginal zone B cells

    Journal: Immunity, Inflammation and Disease

    doi: 10.1002/iid3.134

    The levels of immature B cells decrease dramatically at 17 days post Plasmodium chabaudi infection express AID. AID‐GFP mice were infected with P. chabaudi or given a control injection of uninfected RBC and sacrificed at 17 dpi. Spleens were removed and analyzed by flow cytometry. Representative flow cytometry plots demonstrating the gating strategy for each of the B cell subsets are shown. The left panel is differentiating the mature (CD19+, CD93−) and immature (CD19+, CD93+), and CD93 high B cells based on expression of CD19 and CD93. Transitional B cells subsets are gated into T1 (IgM+, CD23−), T2 (IgM+, CD23+), and T3 (IgM−, CD23+) from the immature B cell subset. Germinal center B cells (PNA high , CD38−) and the rest of the mature B cells are further analyzed to determine the follicular (PNA−, CD38+, CD23+, CD21+) and marginal zone (PNA−, CD38+, CD23 int , CD21 high ) B cells. The bottom panel contains representative flow cytometry histograms demonstrating the expression of AID in each B cell subset ( P. chabaudi infected in black, RBC control in gray histograms) ( n = 3 in two independent experiments).
    Figure Legend Snippet: The levels of immature B cells decrease dramatically at 17 days post Plasmodium chabaudi infection express AID. AID‐GFP mice were infected with P. chabaudi or given a control injection of uninfected RBC and sacrificed at 17 dpi. Spleens were removed and analyzed by flow cytometry. Representative flow cytometry plots demonstrating the gating strategy for each of the B cell subsets are shown. The left panel is differentiating the mature (CD19+, CD93−) and immature (CD19+, CD93+), and CD93 high B cells based on expression of CD19 and CD93. Transitional B cells subsets are gated into T1 (IgM+, CD23−), T2 (IgM+, CD23+), and T3 (IgM−, CD23+) from the immature B cell subset. Germinal center B cells (PNA high , CD38−) and the rest of the mature B cells are further analyzed to determine the follicular (PNA−, CD38+, CD23+, CD21+) and marginal zone (PNA−, CD38+, CD23 int , CD21 high ) B cells. The bottom panel contains representative flow cytometry histograms demonstrating the expression of AID in each B cell subset ( P. chabaudi infected in black, RBC control in gray histograms) ( n = 3 in two independent experiments).

    Techniques Used: Infection, Mouse Assay, Injection, Flow Cytometry, Cytometry, Expressing

    Transitional B cells secrete antibody when isolated from Plasmodium chabaudi‐ infected mice, but not control mice. (A) ELISpot plates coated with anti‐mouse IgM and IgG were plated with sorted T1 (CD19+, CD93+, IgM+, CD23−, CD138−) and T2 (CD19+, CD93+, IgM+, CD23+, CD138−) B cells from P. chabaudi‐ infected mice or control NP‐OVA‐injected mice and spots were detected with a combination of anti‐IgM and −IgG antibodies. T1 and T2 B cells from P. chabaudi‐ infected mice produced significantly more spots than cells from control mice ( n = 3–5). (B) T1 B cells from uninfected mice were cultured with or without CpG on ELISpot plates coated with either anti‐IgM or −IgG antibodies and there was no significant difference between the groups for IgM or IgG secreting cells ( n = 2). (C) T2 B cells treated with CpG had higher numbers of IgG secreting cells and IgM secreting cells in response to CpG. * P
    Figure Legend Snippet: Transitional B cells secrete antibody when isolated from Plasmodium chabaudi‐ infected mice, but not control mice. (A) ELISpot plates coated with anti‐mouse IgM and IgG were plated with sorted T1 (CD19+, CD93+, IgM+, CD23−, CD138−) and T2 (CD19+, CD93+, IgM+, CD23+, CD138−) B cells from P. chabaudi‐ infected mice or control NP‐OVA‐injected mice and spots were detected with a combination of anti‐IgM and −IgG antibodies. T1 and T2 B cells from P. chabaudi‐ infected mice produced significantly more spots than cells from control mice ( n = 3–5). (B) T1 B cells from uninfected mice were cultured with or without CpG on ELISpot plates coated with either anti‐IgM or −IgG antibodies and there was no significant difference between the groups for IgM or IgG secreting cells ( n = 2). (C) T2 B cells treated with CpG had higher numbers of IgG secreting cells and IgM secreting cells in response to CpG. * P

    Techniques Used: Isolation, Infection, Mouse Assay, Enzyme-linked Immunospot, Injection, Produced, Cell Culture

    CpG stimulation leads to AID expression in mature, and T1 B cells and CpG+anti‐IgM resulted in the expression of AID in T2 B cells. B cells enriched by negative selection from AID‐GFP mice were cultured in the presence of media alone, CpG, or CpG+anti‐IgM for 3 days. Populations of mature, T1 and T2 B cell subsets were analyzed for their expression of GFP. Representative flow cytometry showing the mature (CD19+CD93−) and immature (CD19+CD93+) B cells on the left panel. Transitional types 1 and 2 B cells were differentiated in the second column by IgM and CD23 expression (T1 = IgM+, CD23− and T2 = IgM+, CD23 +). The right three columns show AID‐GFP expression for the mature B cells (CD19+ CD93−), T1 cells, and T2 cells compared to a cell proliferation dye ( n = 3 in two independent experiments).
    Figure Legend Snippet: CpG stimulation leads to AID expression in mature, and T1 B cells and CpG+anti‐IgM resulted in the expression of AID in T2 B cells. B cells enriched by negative selection from AID‐GFP mice were cultured in the presence of media alone, CpG, or CpG+anti‐IgM for 3 days. Populations of mature, T1 and T2 B cell subsets were analyzed for their expression of GFP. Representative flow cytometry showing the mature (CD19+CD93−) and immature (CD19+CD93+) B cells on the left panel. Transitional types 1 and 2 B cells were differentiated in the second column by IgM and CD23 expression (T1 = IgM+, CD23− and T2 = IgM+, CD23 +). The right three columns show AID‐GFP expression for the mature B cells (CD19+ CD93−), T1 cells, and T2 cells compared to a cell proliferation dye ( n = 3 in two independent experiments).

    Techniques Used: Expressing, Selection, Mouse Assay, Cell Culture, Flow Cytometry, Cytometry

    21) Product Images from "Stat1 and Stat2 but Not Stat3 Arbitrate Contradictory Growth Signals Elicited by Alpha/Beta Interferon in T Lymphocytes"

    Article Title: Stat1 and Stat2 but Not Stat3 Arbitrate Contradictory Growth Signals Elicited by Alpha/Beta Interferon in T Lymphocytes

    Journal:

    doi: 10.1128/MCB.25.13.5456-5465.2005

    Mitogenic action of IFN-α/β is limited to T lymphocytes and NK cells. (A) Splenocytes, NK cells, or serum-starved fibroblasts from STAT1-deficient mice were stimulated with anti-CD3 (2.5 μg/ml), anti-IgM (10 μg/ml), IL-2
    Figure Legend Snippet: Mitogenic action of IFN-α/β is limited to T lymphocytes and NK cells. (A) Splenocytes, NK cells, or serum-starved fibroblasts from STAT1-deficient mice were stimulated with anti-CD3 (2.5 μg/ml), anti-IgM (10 μg/ml), IL-2

    Techniques Used: Mouse Assay

    22) Product Images from "Selective expansion of myeloid and NK cells in humanized mice yields human-like vaccine responses"

    Article Title: Selective expansion of myeloid and NK cells in humanized mice yields human-like vaccine responses

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07478-2

    NFA2-HIS/Flt3LG mice mount YFV-specific cellular and humoral response. a Schematic representation of the NFA2-HIS mice time course infection experiment. b YFV-17D serum viremia in the peripheral blood of NFA2-HIS/Fluc (blue, n = 10) and NFA2-HIS/Flt3LG (red, n = 14) mice over the course of infection. (+) RNA copies per ml were quantified by RT-qPCR. Limit of detection (dotted line) is shown. Horizontal lines represent median viremia at each time point. * p ≤ 0.05, **** p ≤ 0.0001, ns non-significant (Wilcoxon–Mann–Whitney test). c Absolute cell count of peripheral YFV-specific CD8+ T cells (NS4B/A2+) in the blood of NFA2-HIS/Fluc and NFA2-HIS/Flt3LG mice over the course of YFV-17D infection. Cell counts are shown as per 100 μl of total blood. Horizontal lines represent median cell count at each time point ( n = 4–6). * p ≤ 0.05, ** p ≤ 0.01, ns non-significant (Wilcoxon–Mann–Whitney test). d Absolute count of YFV-specific CD8+ T cells (NS4B/A2+) in the spleen of NFA2-HIS/Fluc and NFA2-HIS/Flt3LG mice at day 20 post infection. Negative controls represent one non-infected NFA2-HIS/Flt3LG mouse and two infected NRGF-HIS/Flt3LG mice (that do not express HLA-A2). Horizontal lines represent median cell count ( n = 4–11). ** p ≤ 0.01 (Wilcoxon–Mann–Whitney test). e , f Relative concentration of human anti-YFV-17D IgM ( e ) and IgG ( f ) antibodies in the serum of NFA2-HIS/Fluc (blue, n = 4) and NFA2-HIS/Flt3LG mice (red, n = 4) over a 6-weeks course of infection. ** p ≤ 0.01, *** p ≤ 0.001, ns non-significant (Wilcoxon–Mann–Whitney test). n.a. non applicable as no mice were analyzed at the time of serum collection. g , h Correlation between YFV-17D viremia (black line) and YFV-IgG relative concentration (colored box and whisker) in the serum of NFA2-HIS/Fluc ( g ) and NFA2-HIS/Flt3LG ( h ) over a 6-weeks course of infection ( n = 4 per group). Medians in each box and whisker are connected together by a colored line (blue for NFA2-HIS/Fluc and red for NFA2-HIS/Flt3LG). Viremia limit of detection (dotted line) is shown. n.a. non applicable as no mice in the control group were alive at the time of serum collection. For panels ( e – h ), bounds of box and whiskers represent the min-to-max absorbance value at each time point. Medians are indicated in each box as center line. i Quantification of YFV-neutralizing activity in the serum of NFA2-NRGF-HIS/Flt3LG mice. Serum neutralizing activity is represented as percentage of YFV-17D infection inhibition (% neutralization). Medians with ranges (min-to-max percentage of neutralization) for both serum dilution are shown ( n = 3). A linear regression (red line) of the average neutralization activity is shown and was used to determine the median neutralization titer (50% inhibition, red number on the x -axis)
    Figure Legend Snippet: NFA2-HIS/Flt3LG mice mount YFV-specific cellular and humoral response. a Schematic representation of the NFA2-HIS mice time course infection experiment. b YFV-17D serum viremia in the peripheral blood of NFA2-HIS/Fluc (blue, n = 10) and NFA2-HIS/Flt3LG (red, n = 14) mice over the course of infection. (+) RNA copies per ml were quantified by RT-qPCR. Limit of detection (dotted line) is shown. Horizontal lines represent median viremia at each time point. * p ≤ 0.05, **** p ≤ 0.0001, ns non-significant (Wilcoxon–Mann–Whitney test). c Absolute cell count of peripheral YFV-specific CD8+ T cells (NS4B/A2+) in the blood of NFA2-HIS/Fluc and NFA2-HIS/Flt3LG mice over the course of YFV-17D infection. Cell counts are shown as per 100 μl of total blood. Horizontal lines represent median cell count at each time point ( n = 4–6). * p ≤ 0.05, ** p ≤ 0.01, ns non-significant (Wilcoxon–Mann–Whitney test). d Absolute count of YFV-specific CD8+ T cells (NS4B/A2+) in the spleen of NFA2-HIS/Fluc and NFA2-HIS/Flt3LG mice at day 20 post infection. Negative controls represent one non-infected NFA2-HIS/Flt3LG mouse and two infected NRGF-HIS/Flt3LG mice (that do not express HLA-A2). Horizontal lines represent median cell count ( n = 4–11). ** p ≤ 0.01 (Wilcoxon–Mann–Whitney test). e , f Relative concentration of human anti-YFV-17D IgM ( e ) and IgG ( f ) antibodies in the serum of NFA2-HIS/Fluc (blue, n = 4) and NFA2-HIS/Flt3LG mice (red, n = 4) over a 6-weeks course of infection. ** p ≤ 0.01, *** p ≤ 0.001, ns non-significant (Wilcoxon–Mann–Whitney test). n.a. non applicable as no mice were analyzed at the time of serum collection. g , h Correlation between YFV-17D viremia (black line) and YFV-IgG relative concentration (colored box and whisker) in the serum of NFA2-HIS/Fluc ( g ) and NFA2-HIS/Flt3LG ( h ) over a 6-weeks course of infection ( n = 4 per group). Medians in each box and whisker are connected together by a colored line (blue for NFA2-HIS/Fluc and red for NFA2-HIS/Flt3LG). Viremia limit of detection (dotted line) is shown. n.a. non applicable as no mice in the control group were alive at the time of serum collection. For panels ( e – h ), bounds of box and whiskers represent the min-to-max absorbance value at each time point. Medians are indicated in each box as center line. i Quantification of YFV-neutralizing activity in the serum of NFA2-NRGF-HIS/Flt3LG mice. Serum neutralizing activity is represented as percentage of YFV-17D infection inhibition (% neutralization). Medians with ranges (min-to-max percentage of neutralization) for both serum dilution are shown ( n = 3). A linear regression (red line) of the average neutralization activity is shown and was used to determine the median neutralization titer (50% inhibition, red number on the x -axis)

    Techniques Used: Mouse Assay, Infection, Quantitative RT-PCR, MANN-WHITNEY, Cell Counting, Concentration Assay, Whisker Assay, Activity Assay, Inhibition, Neutralization

    23) Product Images from "Hypomorphic Mutations in the BCR Signalosome Lead to Selective Immunoglobulin M Deficiency and Impaired B-cell Homeostasis"

    Article Title: Hypomorphic Mutations in the BCR Signalosome Lead to Selective Immunoglobulin M Deficiency and Impaired B-cell Homeostasis

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.02984

    Molecular characterization of novel hypomorphic BTK/BLNK mutations. (A) Gene expression of precursor, secreted and membrane IgM of patients and control EBV-LCL quantified by semi-quantitative cDNA-PCR. (B) Protein expression of monomeric and pentameric IgM of patients and control EBV-LCLs by SDS-PAGE or native-PAGE and detected by western blot. (C) BTK/BLNK Mutation analysis of genomic DNA from peripheral blood. Patient A is hemizygote for a c615G > T in BTK. Patient B is compound heterozygous in BLNK for c328C > G, c472G > T. Healthy controls served as wild type control. (D) Schematic depiction of mutation sites and phosphorylation sites in BTK and BLNK.
    Figure Legend Snippet: Molecular characterization of novel hypomorphic BTK/BLNK mutations. (A) Gene expression of precursor, secreted and membrane IgM of patients and control EBV-LCL quantified by semi-quantitative cDNA-PCR. (B) Protein expression of monomeric and pentameric IgM of patients and control EBV-LCLs by SDS-PAGE or native-PAGE and detected by western blot. (C) BTK/BLNK Mutation analysis of genomic DNA from peripheral blood. Patient A is hemizygote for a c615G > T in BTK. Patient B is compound heterozygous in BLNK for c328C > G, c472G > T. Healthy controls served as wild type control. (D) Schematic depiction of mutation sites and phosphorylation sites in BTK and BLNK.

    Techniques Used: Expressing, Polymerase Chain Reaction, SDS Page, Clear Native PAGE, Western Blot, Mutagenesis

    24) Product Images from "RIG-I-like receptor activation by dengue virus drives follicular T helper cell formation and antibody production"

    Article Title: RIG-I-like receptor activation by dengue virus drives follicular T helper cell formation and antibody production

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006738

    DENV infection of DCs induces Bcl-6 + CXCR5 + PD-1 + T FH formation. Flow cytometry analysis of extracellular CXCR5, PD-1 (A,C) and intracellular Bcl-6 (B,D) expression of differentiated T cells after coculture of naive CD4+ T cells with mock-treated DC or DCs infected with DENV for 48h in the absence or presence of DENV replication inhibitor SDM25N. Numbers in zebra plots of (A) indicate percentage of gated cells. Histograms in (B) represent all T cells from mock coculture (grey), DENV coculture (black) or CXCR5 + PD-1 + T cells from DENV coculture (red) as gated in (A). Results in (D) are relative to fluorescent intensity of DENV samples set as 1. (E) IL-21 in supernatant of differentiated T cells as described in (A) was measured by ELISA. (F) IgM and IgG in the supernatant of B cells cocultured for 7 days with differentiated T cells from mock-treated or DENV-infected DC-T cell cocultures was analyzed by ELISA. Data are representative of at least five (A) or four (B) independent experiments with different donors or are collated data (mean ± s.d.) of five (C), four (D), three (F) or two (E) different donors. ** P
    Figure Legend Snippet: DENV infection of DCs induces Bcl-6 + CXCR5 + PD-1 + T FH formation. Flow cytometry analysis of extracellular CXCR5, PD-1 (A,C) and intracellular Bcl-6 (B,D) expression of differentiated T cells after coculture of naive CD4+ T cells with mock-treated DC or DCs infected with DENV for 48h in the absence or presence of DENV replication inhibitor SDM25N. Numbers in zebra plots of (A) indicate percentage of gated cells. Histograms in (B) represent all T cells from mock coculture (grey), DENV coculture (black) or CXCR5 + PD-1 + T cells from DENV coculture (red) as gated in (A). Results in (D) are relative to fluorescent intensity of DENV samples set as 1. (E) IL-21 in supernatant of differentiated T cells as described in (A) was measured by ELISA. (F) IgM and IgG in the supernatant of B cells cocultured for 7 days with differentiated T cells from mock-treated or DENV-infected DC-T cell cocultures was analyzed by ELISA. Data are representative of at least five (A) or four (B) independent experiments with different donors or are collated data (mean ± s.d.) of five (C), four (D), three (F) or two (E) different donors. ** P

    Techniques Used: Infection, Flow Cytometry, Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

    RLR activation by DENV or synthetic ligands drives T FH formation via IL-27. Flow cytometry analysis of extracellular CXCR5, PD-1 (A,F) and intracellular Bcl-6 (B,E) expression of differentiated T cells resulting from cocultured naive CD4+ T cells with mock-treated DC or DCs infected with DENV (A,B) or stimulated with LPS, poly(I:C)LyoVec or 5’pppRNA-LyoVec (E,F) for 48h in the absence or presence of neutralizing antibodies against IL-27 (F) or after silencing MAVS (A,B). (C) IL-27p28 and IL-27EBI3 mRNA levels in DCs stimulated with poly(I:C)LyoVec or 5’pppRNA-LyoVec for 8h were analyzed using real-time PCR, normalized to GAPDH and set at 1 in 1.0 μg ml -1 stimulated poly(I:C)LyoVec samples. (D) IL-27 in the supernatant of untreated, poly(I:C)LyoVec or 5’pppRNA-LyoVec stimulated DCs after 24h or 48h was analyzed by ELISA. (G) IgM and IgG in the supernatant of B cells cocultured for 7 days with differentiated T cells from cocultures of naive CD4+ T cells with untreated, LPS, poly(I:C)LyoVec or 5’pppRNA-LyoVec stimulated DCs was analyzed by ELISA. Data are representative of at least three (D,E) or two (B) independent experiments with different donors (mean ± s.d. of duplicates in, D) or are collated data (mean ± s.d.) of three (G), four (C,F) or two (A) different donors. ** P
    Figure Legend Snippet: RLR activation by DENV or synthetic ligands drives T FH formation via IL-27. Flow cytometry analysis of extracellular CXCR5, PD-1 (A,F) and intracellular Bcl-6 (B,E) expression of differentiated T cells resulting from cocultured naive CD4+ T cells with mock-treated DC or DCs infected with DENV (A,B) or stimulated with LPS, poly(I:C)LyoVec or 5’pppRNA-LyoVec (E,F) for 48h in the absence or presence of neutralizing antibodies against IL-27 (F) or after silencing MAVS (A,B). (C) IL-27p28 and IL-27EBI3 mRNA levels in DCs stimulated with poly(I:C)LyoVec or 5’pppRNA-LyoVec for 8h were analyzed using real-time PCR, normalized to GAPDH and set at 1 in 1.0 μg ml -1 stimulated poly(I:C)LyoVec samples. (D) IL-27 in the supernatant of untreated, poly(I:C)LyoVec or 5’pppRNA-LyoVec stimulated DCs after 24h or 48h was analyzed by ELISA. (G) IgM and IgG in the supernatant of B cells cocultured for 7 days with differentiated T cells from cocultures of naive CD4+ T cells with untreated, LPS, poly(I:C)LyoVec or 5’pppRNA-LyoVec stimulated DCs was analyzed by ELISA. Data are representative of at least three (D,E) or two (B) independent experiments with different donors (mean ± s.d. of duplicates in, D) or are collated data (mean ± s.d.) of three (G), four (C,F) or two (A) different donors. ** P

    Techniques Used: Activation Assay, Flow Cytometry, Cytometry, Expressing, Infection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    25) Product Images from "Leptin and EGF Supplementation Enhance the Immune System Maturation in Preterm Suckling Rats"

    Article Title: Leptin and EGF Supplementation Enhance the Immune System Maturation in Preterm Suckling Rats

    Journal: Nutrients

    doi: 10.3390/nu11102380

    Plasma Ig concentrations at day 10 and 17. Plasma IgA ( A ), IgM ( B ), and IgG ( C ) concentrations from the four groups: Term (T), Preterm (P), P+Leptin, and P+epidermal growth factor (P+EGF), are expressed as mean ± SD ( n = 8–12 pups/group). Statistical differences: * p
    Figure Legend Snippet: Plasma Ig concentrations at day 10 and 17. Plasma IgA ( A ), IgM ( B ), and IgG ( C ) concentrations from the four groups: Term (T), Preterm (P), P+Leptin, and P+epidermal growth factor (P+EGF), are expressed as mean ± SD ( n = 8–12 pups/group). Statistical differences: * p

    Techniques Used:

    26) Product Images from "A Robust and Versatile Automated Glycoanalytical Technology for Serum Antibodies and Acute Phase Proteins: Ovarian Cancer Case Study *"

    Article Title: A Robust and Versatile Automated Glycoanalytical Technology for Serum Antibodies and Acute Phase Proteins: Ovarian Cancer Case Study *

    Journal: Molecular & Cellular Proteomics : MCP

    doi: 10.1074/mcp.RA119.001531

    Glycosylation traits for selected antibodies (IgG, IgM, IgA) and acute phase proteins (Trf, Hpt, A1AT) for healthy human serum are presented (5 A ). . Glycosylation features of antibodies (IgG, IgM and IgA) and acute phase proteins (Trf, Hpt, A1AT) are presented as control charts (5 B and 5 C ).
    Figure Legend Snippet: Glycosylation traits for selected antibodies (IgG, IgM, IgA) and acute phase proteins (Trf, Hpt, A1AT) for healthy human serum are presented (5 A ). . Glycosylation features of antibodies (IgG, IgM and IgA) and acute phase proteins (Trf, Hpt, A1AT) are presented as control charts (5 B and 5 C ).

    Techniques Used:

    UPLC-HILIC-FLD chromatograms of AQC labeled N -glycans released from human serum for affinity purified glycoproteins IgG, IgM and IgA, Trf, Hpt, A1AT and 2-AB labeled N -glycans released from total serum. Only major glycans are annotated. All major N -glycans identified within the total serum UPLC chromatogram are present in the individual glycoprotein chromatograms which showcases that serum glycosylation is largely dominated by the highest abundance proteins (IgG, IgM and IgA, Trf, Hpt and A1AT). SNFG glycan nomenclature is used throughout.
    Figure Legend Snippet: UPLC-HILIC-FLD chromatograms of AQC labeled N -glycans released from human serum for affinity purified glycoproteins IgG, IgM and IgA, Trf, Hpt, A1AT and 2-AB labeled N -glycans released from total serum. Only major glycans are annotated. All major N -glycans identified within the total serum UPLC chromatogram are present in the individual glycoprotein chromatograms which showcases that serum glycosylation is largely dominated by the highest abundance proteins (IgG, IgM and IgA, Trf, Hpt and A1AT). SNFG glycan nomenclature is used throughout.

    Techniques Used: Hydrophilic Interaction Liquid Chromatography, Labeling, Affinity Purification

    Multiplexed automated serial capture of glycoprotein N -glycoprofiling. 96-well format robotic platform ( A ), specific anti-glycoprotein capture resin packed in PhyNexus phytip ( B ), serial capture of selected glycoproteins 1.Trf, 2.IgG, 3.IgM, 4.IgA, 5.Hpt, 6.A1AT ( C ), 1D SDS-PAGE separation of multiplexed automated capture of six selected glycoproteins from pooled human serum using PhyNexus Phytips. Lane 1: Protein Marker, Lane 2: Trf Standard, Lane 3: Bound Trf, Lane 4: Bound IgG, Lane 5: Bound IgM, Lane 6: Bound IgA, Lane 7: A1AT Standard, Lane 8: Bound A1AT, Lane 9: Hpt Standard, Lane 10: Bound Hpt. The protein bands marked with arrows are traces of albumin protein (non-glycosylated) ( D ), automated glycoprotein sample preparation, PNGaseF release and aminoquinoline carbamate (AQC) labeling of N -glycans ( E ) and finally ultra-high performance liquid chromatography (UPLC) separation and glycan structural analysis ( F ).
    Figure Legend Snippet: Multiplexed automated serial capture of glycoprotein N -glycoprofiling. 96-well format robotic platform ( A ), specific anti-glycoprotein capture resin packed in PhyNexus phytip ( B ), serial capture of selected glycoproteins 1.Trf, 2.IgG, 3.IgM, 4.IgA, 5.Hpt, 6.A1AT ( C ), 1D SDS-PAGE separation of multiplexed automated capture of six selected glycoproteins from pooled human serum using PhyNexus Phytips. Lane 1: Protein Marker, Lane 2: Trf Standard, Lane 3: Bound Trf, Lane 4: Bound IgG, Lane 5: Bound IgM, Lane 6: Bound IgA, Lane 7: A1AT Standard, Lane 8: Bound A1AT, Lane 9: Hpt Standard, Lane 10: Bound Hpt. The protein bands marked with arrows are traces of albumin protein (non-glycosylated) ( D ), automated glycoprotein sample preparation, PNGaseF release and aminoquinoline carbamate (AQC) labeling of N -glycans ( E ) and finally ultra-high performance liquid chromatography (UPLC) separation and glycan structural analysis ( F ).

    Techniques Used: SDS Page, Marker, Sample Prep, Labeling, High Performance Liquid Chromatography

    Serial capture and N -glycoprofiling of human serum IgG, IgM and IgA, purified from human serum. The 25 IgG glycan peak areas (G1-G25), 24 IgM peak areas (M1–24) and 25 IgA glycan peak areas (A1-A25) plotted for five technical replicates over three different days (3 A ). The standard error shown as error bars. Sequencing of AQC labeled IgG, IgM and IgA N -glycans visualized by UPLC-HILIC chromatograms using exoglycosidase enzymes with glucose units (GU) to facilitate glycan identification (3 B ). Digestion of AQC labeled IgG, IgM and IgA N -glycans with addition of sialidase (ABS), galactosidase (BTG), hexosaminidase (GUH), fucosidase (BKF) and mannosidase (JBM) in the following order ABS, ABS+BTG, ABS+BTG+GUH, ABS+BTG+GUH+BKF and ABS+BTG+GUH+BKF+JBM. For IgG, arrows indicate the cleavage of sugar residues for selected peaks: major glycans FA2G2S1 and FA2G2S2. For IgM, arrows indicate the cleavage of sugar residues for selected peaks: major glycans FA2G2S1 and FA2BG2S1. For IgA, arrows indicate the cleavage of sugar residues for selected peaks: major glycans A2G2S1, FA2G2S2 and FA2BG2S2. SNFG nomenclature is used for glycan representation.
    Figure Legend Snippet: Serial capture and N -glycoprofiling of human serum IgG, IgM and IgA, purified from human serum. The 25 IgG glycan peak areas (G1-G25), 24 IgM peak areas (M1–24) and 25 IgA glycan peak areas (A1-A25) plotted for five technical replicates over three different days (3 A ). The standard error shown as error bars. Sequencing of AQC labeled IgG, IgM and IgA N -glycans visualized by UPLC-HILIC chromatograms using exoglycosidase enzymes with glucose units (GU) to facilitate glycan identification (3 B ). Digestion of AQC labeled IgG, IgM and IgA N -glycans with addition of sialidase (ABS), galactosidase (BTG), hexosaminidase (GUH), fucosidase (BKF) and mannosidase (JBM) in the following order ABS, ABS+BTG, ABS+BTG+GUH, ABS+BTG+GUH+BKF and ABS+BTG+GUH+BKF+JBM. For IgG, arrows indicate the cleavage of sugar residues for selected peaks: major glycans FA2G2S1 and FA2G2S2. For IgM, arrows indicate the cleavage of sugar residues for selected peaks: major glycans FA2G2S1 and FA2BG2S1. For IgA, arrows indicate the cleavage of sugar residues for selected peaks: major glycans A2G2S1, FA2G2S2 and FA2BG2S2. SNFG nomenclature is used for glycan representation.

    Techniques Used: Purification, Sequencing, Labeling, Hydrophilic Interaction Liquid Chromatography

    27) Product Images from "CD40 Ligand Deficient C57BL/6 Mouse Is a Potential Surrogate Model of Human X-Linked Hyper IgM (X-HIGM) Syndrome for Characterizing Immune Responses against Pathogens"

    Article Title: CD40 Ligand Deficient C57BL/6 Mouse Is a Potential Surrogate Model of Human X-Linked Hyper IgM (X-HIGM) Syndrome for Characterizing Immune Responses against Pathogens

    Journal: BioMed Research International

    doi: 10.1155/2015/679850

    Basal Ig isotypes sera concentrations of WT ( n = 17) and C57-CD40L −/− ( n = 19) mice. (a) Basal IgM, IgG, and IgA concentrations. Ig concentrations were determined by extrapolation from a standard curve. The total mean IgM concentration was similar among mice strains. Total IgG and IgA serum concentrations of C57-CD40L −/− were significantly lower than concentrations of WT mice. (b) Basal IgG subclasses sera concentrations. IgG3 levels were essentially identical between the two groups and all other subclasses concentrations were significantly higher in WT than in CD40L deficient mice. Mean comparisons were done by Mann-Whitney U -test. WT = wild type.
    Figure Legend Snippet: Basal Ig isotypes sera concentrations of WT ( n = 17) and C57-CD40L −/− ( n = 19) mice. (a) Basal IgM, IgG, and IgA concentrations. Ig concentrations were determined by extrapolation from a standard curve. The total mean IgM concentration was similar among mice strains. Total IgG and IgA serum concentrations of C57-CD40L −/− were significantly lower than concentrations of WT mice. (b) Basal IgG subclasses sera concentrations. IgG3 levels were essentially identical between the two groups and all other subclasses concentrations were significantly higher in WT than in CD40L deficient mice. Mean comparisons were done by Mann-Whitney U -test. WT = wild type.

    Techniques Used: Mouse Assay, Concentration Assay, MANN-WHITNEY

    C. rodentium -specific sera antibodies of wild type (WT) ( n = 6) and C57-CD40L −/− ( n = 6) mice. Mice orally inoculated with 1 × 10 7 CFU of C. rodentium were bled at day 14 after inoculation. Each point of these curves represents the mean ± SD of the OD determinations. (a) The specific IgM titres were essentially identical between the two groups. ((b) and (c)) The specific IgG and IgG2b titres were significantly lower for C57-CD40L −/− mice than WT mice. (d) IgG2c specific antibodies of both WT and C57-CD40L −/− mice were just above baseline and IgG1 and IgG3 isotypes were undetectable. Antibody titres are presented as means and data was analysed by Mann-Whitney U -test. OD = optical density, (1 : n ) = dilution factor.
    Figure Legend Snippet: C. rodentium -specific sera antibodies of wild type (WT) ( n = 6) and C57-CD40L −/− ( n = 6) mice. Mice orally inoculated with 1 × 10 7 CFU of C. rodentium were bled at day 14 after inoculation. Each point of these curves represents the mean ± SD of the OD determinations. (a) The specific IgM titres were essentially identical between the two groups. ((b) and (c)) The specific IgG and IgG2b titres were significantly lower for C57-CD40L −/− mice than WT mice. (d) IgG2c specific antibodies of both WT and C57-CD40L −/− mice were just above baseline and IgG1 and IgG3 isotypes were undetectable. Antibody titres are presented as means and data was analysed by Mann-Whitney U -test. OD = optical density, (1 : n ) = dilution factor.

    Techniques Used: Mouse Assay, MANN-WHITNEY

    28) Product Images from "Association of Anti–Topoisomerase I Antibodies of the IgM Isotype With Disease Progression in Anti–Topoisomerase I–Positive Systemic Sclerosis"

    Article Title: Association of Anti–Topoisomerase I Antibodies of the IgM Isotype With Disease Progression in Anti–Topoisomerase I–Positive Systemic Sclerosis

    Journal: Arthritis & Rheumatology (Hoboken, N.j.)

    doi: 10.1002/art.41403

    Correlation between baseline levels of anti–topoisomerase I antibody (ATA) IgG, IgM, and IgA and modified Rodnan skin thickness score (MRSS) in patients from the Leiden Combined Care in Systemic Sclerosis cohort (n = 103). Spearman’s correlation analyses indicated that only anti–topoisomerase I IgG levels were significantly correlated with MRSS scores.
    Figure Legend Snippet: Correlation between baseline levels of anti–topoisomerase I antibody (ATA) IgG, IgM, and IgA and modified Rodnan skin thickness score (MRSS) in patients from the Leiden Combined Care in Systemic Sclerosis cohort (n = 103). Spearman’s correlation analyses indicated that only anti–topoisomerase I IgG levels were significantly correlated with MRSS scores.

    Techniques Used: Modification

    29) Product Images from "Transient BAFF blockade inhibits type 1 diabetes development in NOD mice by enriching immunoregulatory B-lymphocytes sensitive to deletion by anti-CD20 co-therapy BAFF blockade inhibits type 1 diabetes development in NOD mice by enriching immunoregulatory B-lymphocytes sensitive to deletion by anti-CD20 co-therapy"

    Article Title: Transient BAFF blockade inhibits type 1 diabetes development in NOD mice by enriching immunoregulatory B-lymphocytes sensitive to deletion by anti-CD20 co-therapy BAFF blockade inhibits type 1 diabetes development in NOD mice by enriching immunoregulatory B-lymphocytes sensitive to deletion by anti-CD20 co-therapy

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1700822

    Short-term BAFFR-Fc mediated B-lymphocyte depletion elicits prolonged T1D protection and remaining B-lymphocytes are regulatory in nature A) NOD female mice treated with BAFFR-Fc alone or combination with anti-CD20 between 10–14 weeks of age were assessed for diabetes development. Diabetes development in each of these experimental groups was statistically compared to NOD females treated with ctrl mAb from 10–14 weeks of age (Mantel-Cox analysis). B) Representative flow cytometric plot showing frequency of B 10 cells among CD19 + cells in spleens of NOD mice treated with ctrl mAb, BAFFR-Fc, or anti-CD20 from 10–14 weeks of age (left panel). Summary of the percentage of B 10 cells in the spleens and islets (right panel). Data pooled from three and two independent experiments for B 10 cells in the spleens and islets respectively. C) 1.0×10 5 sort-purified splenic B-lymphocytes from ctrl mAb or BAFFR-Fc-treated NOD mice were cultured for 3 days with or without LPS or anti-IgM plus anti-CD40 and culture supernatant was subsequently measured by ELISA for IL-10. Data pooled from two independent experiments. D) Frequency of B 10 cells amongst CD80 + PD-L2 + or CD80 − PD-L2 − B-lymphocytes in spleens of NOD mice treated with ctrl mAb or BAFFR-Fc from 10–14 weeks of age. E-F) Number of B1a, B1b, B2 B-lymphocytes in the peritoneal cavity (E) and percentage of B 10 cells within the B1a, B1b, B2 B-lymphocytes subsets (F) in NOD female mice treated with ctrl mAb or BAFFR-Fc from 10–14 weeks of age. G) 1.0×10 5 CD25 − CD4 + T-cells from 10 week-old NOD mice were labeled with Cell Proliferation Dye eFluor450 (CPD) and then stimulated with 5 μg/mL plate bound anti-CD3ε and co-cultured for 3 days with 1.0×10 5 sorted splenic B-lymphocytes from ctrl mAb or BAFFR-Fc-treated NOD mice with or without 10 μg/mL LPS stimulation (left panel). Quantification of percent T-cell proliferation suppression (right panel) (n=4 per group). Data are representative of two experiments. H) NOD female mice treated with BAFFR-Fc or ctrl mAb between 10–14 weeks of age were assessed for diabetes development. Upon cessation of transient BAFFR-Fc monotherapy a separate cohort of NOD females subsequently received twice-weekly injections of anti-IL-10 while being monitored for diabetes development. Pair-wise comparisons of diabetes development between each group were calculated using Mantel-Cox analysis. I) Representative flow cytometric plots showing islet infiltrating B-lymphocytes expressing CD73 among live CD19 + B220 + cells from NOD mice treated with ctrl mAb, BAFFR-Fc, or anti-CD20 from 10–14 weeks of age (left panel). Percent CD73 + B-lymphocytes from spleens, PLNs, or islets of NOD mice treated with ctrl mAb, BAFFR-Fc, or anti-CD20 from 10–14 weeks of age (right panel). Data pooled from two independent experiments. All p -values were calculated using Mann-Whitney analyses. Each symbol represents an individual mouse except for BAFFR-Fc group in C and D where each symbol represents sorted B-lymphocytes pooled from 2 or 3 mice; small horizontal lines indicate the Mean±SEM.
    Figure Legend Snippet: Short-term BAFFR-Fc mediated B-lymphocyte depletion elicits prolonged T1D protection and remaining B-lymphocytes are regulatory in nature A) NOD female mice treated with BAFFR-Fc alone or combination with anti-CD20 between 10–14 weeks of age were assessed for diabetes development. Diabetes development in each of these experimental groups was statistically compared to NOD females treated with ctrl mAb from 10–14 weeks of age (Mantel-Cox analysis). B) Representative flow cytometric plot showing frequency of B 10 cells among CD19 + cells in spleens of NOD mice treated with ctrl mAb, BAFFR-Fc, or anti-CD20 from 10–14 weeks of age (left panel). Summary of the percentage of B 10 cells in the spleens and islets (right panel). Data pooled from three and two independent experiments for B 10 cells in the spleens and islets respectively. C) 1.0×10 5 sort-purified splenic B-lymphocytes from ctrl mAb or BAFFR-Fc-treated NOD mice were cultured for 3 days with or without LPS or anti-IgM plus anti-CD40 and culture supernatant was subsequently measured by ELISA for IL-10. Data pooled from two independent experiments. D) Frequency of B 10 cells amongst CD80 + PD-L2 + or CD80 − PD-L2 − B-lymphocytes in spleens of NOD mice treated with ctrl mAb or BAFFR-Fc from 10–14 weeks of age. E-F) Number of B1a, B1b, B2 B-lymphocytes in the peritoneal cavity (E) and percentage of B 10 cells within the B1a, B1b, B2 B-lymphocytes subsets (F) in NOD female mice treated with ctrl mAb or BAFFR-Fc from 10–14 weeks of age. G) 1.0×10 5 CD25 − CD4 + T-cells from 10 week-old NOD mice were labeled with Cell Proliferation Dye eFluor450 (CPD) and then stimulated with 5 μg/mL plate bound anti-CD3ε and co-cultured for 3 days with 1.0×10 5 sorted splenic B-lymphocytes from ctrl mAb or BAFFR-Fc-treated NOD mice with or without 10 μg/mL LPS stimulation (left panel). Quantification of percent T-cell proliferation suppression (right panel) (n=4 per group). Data are representative of two experiments. H) NOD female mice treated with BAFFR-Fc or ctrl mAb between 10–14 weeks of age were assessed for diabetes development. Upon cessation of transient BAFFR-Fc monotherapy a separate cohort of NOD females subsequently received twice-weekly injections of anti-IL-10 while being monitored for diabetes development. Pair-wise comparisons of diabetes development between each group were calculated using Mantel-Cox analysis. I) Representative flow cytometric plots showing islet infiltrating B-lymphocytes expressing CD73 among live CD19 + B220 + cells from NOD mice treated with ctrl mAb, BAFFR-Fc, or anti-CD20 from 10–14 weeks of age (left panel). Percent CD73 + B-lymphocytes from spleens, PLNs, or islets of NOD mice treated with ctrl mAb, BAFFR-Fc, or anti-CD20 from 10–14 weeks of age (right panel). Data pooled from two independent experiments. All p -values were calculated using Mann-Whitney analyses. Each symbol represents an individual mouse except for BAFFR-Fc group in C and D where each symbol represents sorted B-lymphocytes pooled from 2 or 3 mice; small horizontal lines indicate the Mean±SEM.

    Techniques Used: Mouse Assay, Flow Cytometry, Purification, Cell Culture, Enzyme-linked Immunosorbent Assay, Labeling, Expressing, MANN-WHITNEY

    30) Product Images from "DYRK1A controls the transition from proliferation to quiescence during lymphoid development by destabilizing Cyclin D3"

    Article Title: DYRK1A controls the transition from proliferation to quiescence during lymphoid development by destabilizing Cyclin D3

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20150002

    Dyrk1a -deficient quiescent DP thymocytes and small pre–B cells fail to repress E2F target gene transcription . (A and B) Up- and down-regulated transcripts were identified by RNA-sequencing of FACS-purified quiescent DP thymocytes (CD4 + /CD8 + /DNA content 2N /Pyronin Y low ), cycling DP thymocytes (CD4 + /CD8 + /DNA content > 2N /Pyronin Y high ), small pre–B cells (IgM − /B220 + /CD43 − /FSC low ), large pre–B cells (IgM − /B220 + /CD43 low /FSC high ), and granulocytes (Gr-1 high /Mac-1/CD11b high ) from Dyrk1a f/f Mx1-Cre − (Control) and Dyrk1a f/f Mx1-Cre + (CKO) mice. Venn diagrams depict shared up-regulated (A) and down-regulated (B) transcripts identified by RNA-sequencing in the three quiescent cell populations. (C–F) mRNA expression of E2F target genes was assessed by qRT-PCR in FACS-purified small pre–B cells (C), quiescent DP thymocytes (D), large pre–B cells (E), and cycling DP thymocytes (F) from Control and CKO mice 2 wk after pI:pC treatment. Transcript levels were normalized to Actb expression. PCRs were performed using 3 independently sorted pairs of samples (each with 1 mouse per genotype) for pre–B cells, and pooled samples from 3 mice per genotype for thymocytes. Error bars depict SD of triplicate wells for representative samples. *, P
    Figure Legend Snippet: Dyrk1a -deficient quiescent DP thymocytes and small pre–B cells fail to repress E2F target gene transcription . (A and B) Up- and down-regulated transcripts were identified by RNA-sequencing of FACS-purified quiescent DP thymocytes (CD4 + /CD8 + /DNA content 2N /Pyronin Y low ), cycling DP thymocytes (CD4 + /CD8 + /DNA content > 2N /Pyronin Y high ), small pre–B cells (IgM − /B220 + /CD43 − /FSC low ), large pre–B cells (IgM − /B220 + /CD43 low /FSC high ), and granulocytes (Gr-1 high /Mac-1/CD11b high ) from Dyrk1a f/f Mx1-Cre − (Control) and Dyrk1a f/f Mx1-Cre + (CKO) mice. Venn diagrams depict shared up-regulated (A) and down-regulated (B) transcripts identified by RNA-sequencing in the three quiescent cell populations. (C–F) mRNA expression of E2F target genes was assessed by qRT-PCR in FACS-purified small pre–B cells (C), quiescent DP thymocytes (D), large pre–B cells (E), and cycling DP thymocytes (F) from Control and CKO mice 2 wk after pI:pC treatment. Transcript levels were normalized to Actb expression. PCRs were performed using 3 independently sorted pairs of samples (each with 1 mouse per genotype) for pre–B cells, and pooled samples from 3 mice per genotype for thymocytes. Error bars depict SD of triplicate wells for representative samples. *, P

    Techniques Used: RNA Sequencing Assay, FACS, Purification, Mouse Assay, Expressing, Quantitative RT-PCR

    31) Product Images from "Perivascular Adipose Tissue Harbors Atheroprotective IgM-Producing B Cells"

    Article Title: Perivascular Adipose Tissue Harbors Atheroprotective IgM-Producing B Cells

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2017.00719

    B-1b cells and IgM secreting B cells are increased in the PVAT of mice with B cell specific deletion of Id3 (ApoE −/− Id3 BKO ). Flow cytometry quantification in the PVAT of 8 weeks old ApoE −/− Id3 WT ( n = 11) and ApoE −/− Id3 BKO ( n = 12) mice for (A) total CD19 + B cells, (B) B-1 and B-2 cells, (C) B-1/B-2 ratio, (D) B-1a and B-1b cells, and (E) ELISPOT for total IgM secreting cells. (F) Percentage of malondialdehyde modified low density lipoprotein (MDA-LDL) specific IgM secreting cells in spleen, BM and PVAT of ApoE −/− Id3 WT ( n = 5) and ApoE −/− Id3 BKO ( n = 4–5) mice as measured by ELISPOT. Results are mean ± SEM, unpaired student t -test was performed ( * p
    Figure Legend Snippet: B-1b cells and IgM secreting B cells are increased in the PVAT of mice with B cell specific deletion of Id3 (ApoE −/− Id3 BKO ). Flow cytometry quantification in the PVAT of 8 weeks old ApoE −/− Id3 WT ( n = 11) and ApoE −/− Id3 BKO ( n = 12) mice for (A) total CD19 + B cells, (B) B-1 and B-2 cells, (C) B-1/B-2 ratio, (D) B-1a and B-1b cells, and (E) ELISPOT for total IgM secreting cells. (F) Percentage of malondialdehyde modified low density lipoprotein (MDA-LDL) specific IgM secreting cells in spleen, BM and PVAT of ApoE −/− Id3 WT ( n = 5) and ApoE −/− Id3 BKO ( n = 4–5) mice as measured by ELISPOT. Results are mean ± SEM, unpaired student t -test was performed ( * p

    Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunospot, Modification, Multiple Displacement Amplification

    IgM secreting B-1 cells reside in PVAT. (A) Gating strategy of B-1 (CD19 + , B220 lo- mid ) and B2 (CD19 + , B220 hi ) cells. B-1 (IgD − CD43 + ) and B-2 (IgD + CD43 − ) cells were further confirmed based on surface expression of IgD and CD43. (B) Quantification of total numbers of B-1 and B-2 cells in the aorta and PVAT. (C) Comparative ratio of B-1 to B-2 cells in spleen, bone marrow (BM) and PVAT. (D) B-1a and B-1b cells were gated from total B-1 cells and absolute numbers were quantified in aorta and PVAT of young ApoE −/− mice. (E) IgM antibody production was measured by ELISPOT in aorta and PVAT of Chow diet fed young ApoE −/− mice ( n = 7) (representative plate and quantitation). Results are mean ± SEM, unpaired student t -test was performed. Repeated measures one way ANOVA with Bonferroni's multiple comparison post-test was used to compare multiple groups ( * p
    Figure Legend Snippet: IgM secreting B-1 cells reside in PVAT. (A) Gating strategy of B-1 (CD19 + , B220 lo- mid ) and B2 (CD19 + , B220 hi ) cells. B-1 (IgD − CD43 + ) and B-2 (IgD + CD43 − ) cells were further confirmed based on surface expression of IgD and CD43. (B) Quantification of total numbers of B-1 and B-2 cells in the aorta and PVAT. (C) Comparative ratio of B-1 to B-2 cells in spleen, bone marrow (BM) and PVAT. (D) B-1a and B-1b cells were gated from total B-1 cells and absolute numbers were quantified in aorta and PVAT of young ApoE −/− mice. (E) IgM antibody production was measured by ELISPOT in aorta and PVAT of Chow diet fed young ApoE −/− mice ( n = 7) (representative plate and quantitation). Results are mean ± SEM, unpaired student t -test was performed. Repeated measures one way ANOVA with Bonferroni's multiple comparison post-test was used to compare multiple groups ( * p

    Techniques Used: Expressing, Mouse Assay, Enzyme-linked Immunospot, Quantitation Assay

    32) Product Images from "Heparosan-coated liposomes for drug delivery"

    Article Title: Heparosan-coated liposomes for drug delivery

    Journal: Glycobiology

    doi: 10.1093/glycob/cwx070

    Immunological challenge in rats with HEP-DiPalm and ELISA assessment. A set of three rats ( #1, 2, 3 ) then tested for the induction of an IgM or IgG response by ELISA (representative set of assays with averaged triplicate wells with standard deviation is presented). Overall, the signal from sera of rats before and after injection with HEP-DiPalm micelles (prelipid, white ; postlipid, black ) tested in the control wells coated with bovine serum albumin ( BSA ) was equivalent to the wells coated with HEP-BSA ( HEP ) or HEP-DiPalm ( DiPalm ) indicating that HEP is not significantly immunogenic, therefore, should be useful in multidose or long-term therapeutics.
    Figure Legend Snippet: Immunological challenge in rats with HEP-DiPalm and ELISA assessment. A set of three rats ( #1, 2, 3 ) then tested for the induction of an IgM or IgG response by ELISA (representative set of assays with averaged triplicate wells with standard deviation is presented). Overall, the signal from sera of rats before and after injection with HEP-DiPalm micelles (prelipid, white ; postlipid, black ) tested in the control wells coated with bovine serum albumin ( BSA ) was equivalent to the wells coated with HEP-BSA ( HEP ) or HEP-DiPalm ( DiPalm ) indicating that HEP is not significantly immunogenic, therefore, should be useful in multidose or long-term therapeutics.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Standard Deviation, Injection

    Representation of HEP-liposome assembly. Models of HEP-coated liposome assemblies with 13.3-kDa ( top ) and 7-kDa ( bottom ) HEP-lipid, based on typical molecular configurations from equilibrated coarse-grained simulations. The liposome and sugars are represented graphically by equivalent spheres (roughly to scale), and part of the liposome has been cut-away for viewing purposes. Potential interaction proteins are depicted (human serum albumin, HSA ; immunoglobulins IgG or IgM ; middle ), drawn to exact scale using coordinates from the Protein Data Bank. Less surface area is available for direct protein binding to liposomes coated with 0.5 mol% 13.3-kDa HEP-lipid compared to 0.5 mol% 7-kDa HEP-lipid, suggesting increased liposomal protection from clearance by the immune system. The image was rendered using POV-Ray software.
    Figure Legend Snippet: Representation of HEP-liposome assembly. Models of HEP-coated liposome assemblies with 13.3-kDa ( top ) and 7-kDa ( bottom ) HEP-lipid, based on typical molecular configurations from equilibrated coarse-grained simulations. The liposome and sugars are represented graphically by equivalent spheres (roughly to scale), and part of the liposome has been cut-away for viewing purposes. Potential interaction proteins are depicted (human serum albumin, HSA ; immunoglobulins IgG or IgM ; middle ), drawn to exact scale using coordinates from the Protein Data Bank. Less surface area is available for direct protein binding to liposomes coated with 0.5 mol% 13.3-kDa HEP-lipid compared to 0.5 mol% 7-kDa HEP-lipid, suggesting increased liposomal protection from clearance by the immune system. The image was rendered using POV-Ray software.

    Techniques Used: Protein Binding, Software

    33) Product Images from "Disease-Specific Autoantibodies Induce Trained Immunity in RA Synovial Tissues and Its Gene Signature Correlates with the Response to Clinical Therapy"

    Article Title: Disease-Specific Autoantibodies Induce Trained Immunity in RA Synovial Tissues and Its Gene Signature Correlates with the Response to Clinical Therapy

    Journal: Mediators of Inflammation

    doi: 10.1155/2020/2109325

    Enhanced LPS response by ACPA IgG-primed monocytes. (a–d) ELISA quantification of TNF α , IL6, CXCL8, and CCL2 in the supernatant during monocyte priming. After priming, the Mo(cACPA/IgG), Mo(cRF/IgM), Mo(cIgG), Mo(control), and Mo(LPS) were treated with LPS (10 ng/ml); monocytes treated in RPMI 1640 medium were used as the control (RPMI). (e–h) ELISA quantification of TNF α , IL6, CXCL8, and CCL2. Each dot represents one donor ( n = 6). (i) The Mo(cIgG) and Mo(control) were stimulated with LPS (10 ng/ml) for 2 h, followed by intracellular staining of TNF α . (j) Cells stained with isotype control Abs (filled histogram) were used as negative controls. The MFI of TNF α was compared. Data (mean ± SEM) are from three independent experiments. NS: no statistical significance; ∗ P
    Figure Legend Snippet: Enhanced LPS response by ACPA IgG-primed monocytes. (a–d) ELISA quantification of TNF α , IL6, CXCL8, and CCL2 in the supernatant during monocyte priming. After priming, the Mo(cACPA/IgG), Mo(cRF/IgM), Mo(cIgG), Mo(control), and Mo(LPS) were treated with LPS (10 ng/ml); monocytes treated in RPMI 1640 medium were used as the control (RPMI). (e–h) ELISA quantification of TNF α , IL6, CXCL8, and CCL2. Each dot represents one donor ( n = 6). (i) The Mo(cIgG) and Mo(control) were stimulated with LPS (10 ng/ml) for 2 h, followed by intracellular staining of TNF α . (j) Cells stained with isotype control Abs (filled histogram) were used as negative controls. The MFI of TNF α was compared. Data (mean ± SEM) are from three independent experiments. NS: no statistical significance; ∗ P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Staining

    Schematic overview of in vitro trained immunity model by RA autoantibodies. Freshly isolated CD14+ human monocytes were cultured for 24 h in RPMI 1640 medium in Petri dishes precoated with 10 μ g/ml purified ACPA IgG (cACPA/IgG), RF IgM (cRF/IgM), or IVIG (cIgG). Control cells were cultured with RPMI 1640 medium or in the presence of LPS (1 μ g/ml). After priming, the monocytes were detached, washed, and subcultured in 96-well plates for resting. Then, the Mo(cACPA/IgG), Mo(cRF/IgM), Mo(cIgG), Mo(control), and Mo(LPS) were stimulated by LPS (10 ng/ml) for 24 h. Cytokines and chemokines in the supernatants from either the priming or stimulation phase were measured by ELISA.
    Figure Legend Snippet: Schematic overview of in vitro trained immunity model by RA autoantibodies. Freshly isolated CD14+ human monocytes were cultured for 24 h in RPMI 1640 medium in Petri dishes precoated with 10 μ g/ml purified ACPA IgG (cACPA/IgG), RF IgM (cRF/IgM), or IVIG (cIgG). Control cells were cultured with RPMI 1640 medium or in the presence of LPS (1 μ g/ml). After priming, the monocytes were detached, washed, and subcultured in 96-well plates for resting. Then, the Mo(cACPA/IgG), Mo(cRF/IgM), Mo(cIgG), Mo(control), and Mo(LPS) were stimulated by LPS (10 ng/ml) for 24 h. Cytokines and chemokines in the supernatants from either the priming or stimulation phase were measured by ELISA.

    Techniques Used: In Vitro, Isolation, Cell Culture, Purification, Enzyme-linked Immunosorbent Assay

    34) Product Images from "Evaluation of Recombinant Oocyst Protein CP41 for Detection of Cryptosporidium-Specific Antibodies"

    Article Title: Evaluation of Recombinant Oocyst Protein CP41 for Detection of Cryptosporidium-Specific Antibodies

    Journal: Clinical and Diagnostic Laboratory Immunology

    doi: 10.1128/CDLI.12.2.268-272.2005

    Percentages of IgM- and IgG-reactive sera.
    Figure Legend Snippet: Percentages of IgM- and IgG-reactive sera.

    Techniques Used:

    Percentages of IgM- and IgG-reactive sera.
    Figure Legend Snippet: Percentages of IgM- and IgG-reactive sera.

    Techniques Used:

    35) Product Images from "Endophilin A2 regulates B cell protein trafficking and humoral responses"

    Article Title: Endophilin A2 regulates B cell protein trafficking and humoral responses

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.050419

    Endophilin A2 positively regulates BCR internalization but is dispensable for antigen presentation. (A) Soluble anti-IgM internalization in CRISPR-targeted follicular B cells. N = 11-12 mice. (B) Soluble anti-IgD internalization. N = 3 mice. (C) Soluble anti-IgG internalization in GC B cells from SRBC-immunized mice. N = 4 mice. (A-C) Data show mean ± SEM. P, statistical significance from two-way ANOVA. (D) Internalization of anti-IgM from PMS quantified in fixed naïve B cells at 10 min post synapse formation. N =172 and 97 cells analyzed in one out of 2 representative experiments. P = 0.0001 in unpaired t test. (E) Surface IgM or IgD MFI relative to WT in naïve splenic B cells. N = 3 mice. (F) IgM degradation following total cell surface biotinylation, measured as loss of biotin-labelled IgM in cell lysates over time. (G) Western densitometry quantifying IgM degradation assay. One representative experiment of three independent tests. (H) MHCII antigen presentation in SWHEL B cells 24 hours post adoptive transfer and immunization with Ea-HEL-SRBC conjugate as detected by anti-Y-Ae surface stain. (I) Relative anti-Y-Ae MFI in CRISPR-targeted B cells normalized to HEL-only immunized mice. N = 10 immunized mice across 3 experiments. (J) In vitro antigen presentation in CRISPR-targeted B cells measured by anti-Y-Ae. N = 8 independent cultures in 2 experiments. (K) Proliferation (CFSE dilution) of OTII CD4 cells in cocultures with anti-Igκ-OVA pulsed B cells; and quantification from 6-8 mice.
    Figure Legend Snippet: Endophilin A2 positively regulates BCR internalization but is dispensable for antigen presentation. (A) Soluble anti-IgM internalization in CRISPR-targeted follicular B cells. N = 11-12 mice. (B) Soluble anti-IgD internalization. N = 3 mice. (C) Soluble anti-IgG internalization in GC B cells from SRBC-immunized mice. N = 4 mice. (A-C) Data show mean ± SEM. P, statistical significance from two-way ANOVA. (D) Internalization of anti-IgM from PMS quantified in fixed naïve B cells at 10 min post synapse formation. N =172 and 97 cells analyzed in one out of 2 representative experiments. P = 0.0001 in unpaired t test. (E) Surface IgM or IgD MFI relative to WT in naïve splenic B cells. N = 3 mice. (F) IgM degradation following total cell surface biotinylation, measured as loss of biotin-labelled IgM in cell lysates over time. (G) Western densitometry quantifying IgM degradation assay. One representative experiment of three independent tests. (H) MHCII antigen presentation in SWHEL B cells 24 hours post adoptive transfer and immunization with Ea-HEL-SRBC conjugate as detected by anti-Y-Ae surface stain. (I) Relative anti-Y-Ae MFI in CRISPR-targeted B cells normalized to HEL-only immunized mice. N = 10 immunized mice across 3 experiments. (J) In vitro antigen presentation in CRISPR-targeted B cells measured by anti-Y-Ae. N = 8 independent cultures in 2 experiments. (K) Proliferation (CFSE dilution) of OTII CD4 cells in cocultures with anti-Igκ-OVA pulsed B cells; and quantification from 6-8 mice.

    Techniques Used: CRISPR, Mouse Assay, Western Blot, Degradation Assay, Adoptive Transfer Assay, Staining, In Vitro

    36) Product Images from "Induction and Kinetics of Complement-Fixing Antibodies Against Plasmodium vivax Merozoite Surface Protein 3α and Relationship With Immunoglobulin G Subclasses and Immunoglobulin M"

    Article Title: Induction and Kinetics of Complement-Fixing Antibodies Against Plasmodium vivax Merozoite Surface Protein 3α and Relationship With Immunoglobulin G Subclasses and Immunoglobulin M

    Journal: The Journal of Infectious Diseases

    doi: 10.1093/infdis/jiz407

    Functional C1q-fixing capacity of antibody isotypes. A, Correlations matrixes between immunoglobulin G1, G3, and M (IgG1, IgG3, and IgM) and C1q-fixing antibodies to Plasmodium vivax merozoite surface protein 3α (PvMSP3α) in infected children, infected adults, and uninfected adults. Values represent Spearman correlation coefficients, and blue boxes indicate statistical significance ( P
    Figure Legend Snippet: Functional C1q-fixing capacity of antibody isotypes. A, Correlations matrixes between immunoglobulin G1, G3, and M (IgG1, IgG3, and IgM) and C1q-fixing antibodies to Plasmodium vivax merozoite surface protein 3α (PvMSP3α) in infected children, infected adults, and uninfected adults. Values represent Spearman correlation coefficients, and blue boxes indicate statistical significance ( P

    Techniques Used: Functional Assay, Infection

    Seroprevalence and magnitude of immunoglobulin G and M (IgG and IgM) antibodies to Plasmodium vivax merozoite surface protein 3α (PvMSP3α) in children and adults with P. vivax malaria and uninfected adults. A, Seroprevalence of IgG and IgM antibodies against different regions of PvMSP3α. The positive threshold for seroprevalence was calculated as above the mean plus 3 standard deviations of absorbance detected in malaria-naive Australian donors. Numbers atop brackets are P values; the χ 2 test was used for comparisons between groups. B, Cumulative IgG and IgM antibody responses targeting different regions of PvMSP3α. Data represent percentage of individuals who are positive for the protein regions tested. C, Magnitudes of IgG and IgM antibody responses against different regions of PvMSP3α. For boxplots, the lower and upper lines of boxplot represent the first and third quartiles, whisker lines correspond to the highest and lowest values no further than 1.5 interquartile range, and horizontal lines within boxes indicate medians. Data beyond the whisker lines are treated as outliers, represented as circles, squares and triangles. Numbers atop brackets are P values; the Mann-Whitney test was used for comparisons between groups. Abbreviation: OD 450 , optical density at 450 nm.
    Figure Legend Snippet: Seroprevalence and magnitude of immunoglobulin G and M (IgG and IgM) antibodies to Plasmodium vivax merozoite surface protein 3α (PvMSP3α) in children and adults with P. vivax malaria and uninfected adults. A, Seroprevalence of IgG and IgM antibodies against different regions of PvMSP3α. The positive threshold for seroprevalence was calculated as above the mean plus 3 standard deviations of absorbance detected in malaria-naive Australian donors. Numbers atop brackets are P values; the χ 2 test was used for comparisons between groups. B, Cumulative IgG and IgM antibody responses targeting different regions of PvMSP3α. Data represent percentage of individuals who are positive for the protein regions tested. C, Magnitudes of IgG and IgM antibody responses against different regions of PvMSP3α. For boxplots, the lower and upper lines of boxplot represent the first and third quartiles, whisker lines correspond to the highest and lowest values no further than 1.5 interquartile range, and horizontal lines within boxes indicate medians. Data beyond the whisker lines are treated as outliers, represented as circles, squares and triangles. Numbers atop brackets are P values; the Mann-Whitney test was used for comparisons between groups. Abbreviation: OD 450 , optical density at 450 nm.

    Techniques Used: Whisker Assay, MANN-WHITNEY

    Antibody kinetics profile across 28-day follow-up against Plasmodium vivax merozoite surface protein 3α (PvMSP3α). Magnitudes of C1q-fixing antibodies, immunoglobulin G1 G3, and M (IgG1, IgG3, and IgM) against different regions of PvMSP3α were compared between day 0 and days 7 and 28 follow-up. Gray dots and lines represent individual antibody magnitudes over time; horizontal lines, medians. Numbers atop brackets are P values, calculated with the Wilcoxon signed ranked test. Abbreviation: OD 450 , optical density at 450 nm.
    Figure Legend Snippet: Antibody kinetics profile across 28-day follow-up against Plasmodium vivax merozoite surface protein 3α (PvMSP3α). Magnitudes of C1q-fixing antibodies, immunoglobulin G1 G3, and M (IgG1, IgG3, and IgM) against different regions of PvMSP3α were compared between day 0 and days 7 and 28 follow-up. Gray dots and lines represent individual antibody magnitudes over time; horizontal lines, medians. Numbers atop brackets are P values, calculated with the Wilcoxon signed ranked test. Abbreviation: OD 450 , optical density at 450 nm.

    Techniques Used:

    37) Product Images from "Quantification, epitope mapping and genotype cross-reactivity of hepatitis B preS-specific antibodies in subjects vaccinated with different dosage regimens of BM32"

    Article Title: Quantification, epitope mapping and genotype cross-reactivity of hepatitis B preS-specific antibodies in subjects vaccinated with different dosage regimens of BM32

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2020.102953

    Comparison of preS-specific isotype and IgG subclasses responses in the four groups of patients. Shown are optical density (OD) values ( y -axes) corresponding to preS-specific IgG, IgA, IgM, IgE and IgG subclass (IgG 1 –IgG 4 ) levels in patients immunized with 3, 4 or 5 injections of BM32 or with placebo (see inlay) at baseline (V3), after one (V8) or four (V11) months treatment ( x -axes). Medians (horizontal bars) and significant differences are indicated: * p
    Figure Legend Snippet: Comparison of preS-specific isotype and IgG subclasses responses in the four groups of patients. Shown are optical density (OD) values ( y -axes) corresponding to preS-specific IgG, IgA, IgM, IgE and IgG subclass (IgG 1 –IgG 4 ) levels in patients immunized with 3, 4 or 5 injections of BM32 or with placebo (see inlay) at baseline (V3), after one (V8) or four (V11) months treatment ( x -axes). Medians (horizontal bars) and significant differences are indicated: * p

    Techniques Used:

    38) Product Images from "Immunogenicity of Non-Living Anthrax Vaccine Candidates in Cattle and Protective Efficacy of Immune Sera in A/J Mouse Model Compared to the Sterne Live Spore Vaccine"

    Article Title: Immunogenicity of Non-Living Anthrax Vaccine Candidates in Cattle and Protective Efficacy of Immune Sera in A/J Mouse Model Compared to the Sterne Live Spore Vaccine

    Journal: Pathogens

    doi: 10.3390/pathogens9070557

    The anti-FIS IgM and IgG subclasses (IgG1 and IgG2) ELISA titres of cattle are vaccinated at week 0 and 3 with either PrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), CrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), SLSV (n = 8) and Emulsigen-D ® /Alhydrogel ® adjuvants (NegCtl) (n = 4) are presented as box and whisker plots. The cattle sera samples were collected before the vaccinations at week 0 and 3 as well as at week 5 and 12. Sera dilution started at a concentration of 1:50 and values below the (
    Figure Legend Snippet: The anti-FIS IgM and IgG subclasses (IgG1 and IgG2) ELISA titres of cattle are vaccinated at week 0 and 3 with either PrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), CrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), SLSV (n = 8) and Emulsigen-D ® /Alhydrogel ® adjuvants (NegCtl) (n = 4) are presented as box and whisker plots. The cattle sera samples were collected before the vaccinations at week 0 and 3 as well as at week 5 and 12. Sera dilution started at a concentration of 1:50 and values below the (

    Techniques Used: Enzyme-linked Immunosorbent Assay, Whisker Assay, Concentration Assay

    The anti-rPA IgM and IgG subclasses (IgG1 and IgG2) ELISA titres of cattle are vaccinated at week 0 and 3 with either PrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), CrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), SLSV (n = 8) and Emulsigen-D ® /Alhydrogel ® adjuvants (NegCtl) (n = 4) are presented as box and whisker plots. The cattle sera samples were collected before the vaccinations at week 0 and 3 as well as at week 5 and 12. Sera dilution started at a concentration of 1:50 and values below the (
    Figure Legend Snippet: The anti-rPA IgM and IgG subclasses (IgG1 and IgG2) ELISA titres of cattle are vaccinated at week 0 and 3 with either PrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), CrPA+FIS+Emulsigen-D ® /Alhydrogel ® adjuvants (n = 8), SLSV (n = 8) and Emulsigen-D ® /Alhydrogel ® adjuvants (NegCtl) (n = 4) are presented as box and whisker plots. The cattle sera samples were collected before the vaccinations at week 0 and 3 as well as at week 5 and 12. Sera dilution started at a concentration of 1:50 and values below the (

    Techniques Used: Recombinase Polymerase Amplification, Enzyme-linked Immunosorbent Assay, Whisker Assay, Concentration Assay

    39) Product Images from "Ex vivo engineered immune organoids for controlled germinal center reactions"

    Article Title: Ex vivo engineered immune organoids for controlled germinal center reactions

    Journal: Biomaterials

    doi: 10.1016/j.biomaterials.2015.06.002

    Antibody isotype class switching in the immune organoid. (A) Gating strategy for GC phenotype analysis where CD19+ B cells were double stained with IgM and IgGl. Naïve B cells on day 0 predominantly express IgM and no IgG1. (B) Increase in the
    Figure Legend Snippet: Antibody isotype class switching in the immune organoid. (A) Gating strategy for GC phenotype analysis where CD19+ B cells were double stained with IgM and IgGl. Naïve B cells on day 0 predominantly express IgM and no IgG1. (B) Increase in the

    Techniques Used: Staining

    40) Product Images from "Lentiviral Vector Delivery of Human Interleukin-7 (hIL-7) to Human Immune System (HIS) Mice Expands T Lymphocyte Populations"

    Article Title: Lentiviral Vector Delivery of Human Interleukin-7 (hIL-7) to Human Immune System (HIS) Mice Expands T Lymphocyte Populations

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0012009

    Lentiviral vector delivery of hIL-7 improves T cell levels in the spleens and lymph nodes of HIS mice, and increases BCL2 expression. Spleens were removed from hIL-7 (low dose group) or luciferase expressing mice and weighed. b. Splenic sections were H E stained (scale bar = 1 mm). c. Spleens and lymph nodes were processed into single cell suspensions and flow cytometry was used to analyze the percentage of human CD3+ versus CD19+ cells in the different groups. CD3+ cells were analyzed to quantify CD4+ and CD8+ subsets. d. Absolute cell numbers for the indicated lineages were determined using splenocytes from hIL-7 expressing or control mice. e. Serial splenic sections were stained with antibodies against human CD3 (scale bar = 100 µm) or CD20 (scale bar = 100 µm), and another set with CD3 (scale bar = 100 µm) and BCL2 (scale bar = 100 µm). f. Serum from both low and high dose hIL-7 expressing mice or luciferase controls was assayed to determine the concentrations of total IgM or total IgG.
    Figure Legend Snippet: Lentiviral vector delivery of hIL-7 improves T cell levels in the spleens and lymph nodes of HIS mice, and increases BCL2 expression. Spleens were removed from hIL-7 (low dose group) or luciferase expressing mice and weighed. b. Splenic sections were H E stained (scale bar = 1 mm). c. Spleens and lymph nodes were processed into single cell suspensions and flow cytometry was used to analyze the percentage of human CD3+ versus CD19+ cells in the different groups. CD3+ cells were analyzed to quantify CD4+ and CD8+ subsets. d. Absolute cell numbers for the indicated lineages were determined using splenocytes from hIL-7 expressing or control mice. e. Serial splenic sections were stained with antibodies against human CD3 (scale bar = 100 µm) or CD20 (scale bar = 100 µm), and another set with CD3 (scale bar = 100 µm) and BCL2 (scale bar = 100 µm). f. Serum from both low and high dose hIL-7 expressing mice or luciferase controls was assayed to determine the concentrations of total IgM or total IgG.

    Techniques Used: Plasmid Preparation, Mouse Assay, Expressing, Luciferase, Staining, Flow Cytometry, Cytometry

    Related Articles

    Flow Cytometry:

    Article Title: Intact CD100–CD72 Interaction Necessary for TCR-Induced T Cell Proliferation
    Article Snippet: .. Flow Cytometry Fresh or thawed PBMCs were stained for cell surface markers with fluorochrome-conjugated antibodies against the following proteins (dilutions indicated): CD3 1:200 (UCHT1, eBioscience), CD19 1:100 (HIB19, BD Biosciences), CD45 1:100 (HI30, BD Biosciences), CD4 1:100 (RPA-T4, eBioscience), CD8a 1:200 (RPA-T8, eBioscience), CD100 1:50 (A8, Biolegend), CD100 1:50 (unconjugated, 133/1C6, eBioscience), mouse IgM 1:100 (secondary ab for anti-CD100, eB121-15F9, eBioscience), mouse IgG 1:100 (secondary ab for other anti-CD100, Sigma-Aldrich), CD72 1:100 (3F3, Biolegend), and dead cell marker (DCM) 1:200 (Zombie NIR™ Fixable Viability Kit, Biolegend). .. Data were acquired on a FACSVerse™ flow cytometer (BD Biosciences) or on a LSRFortessa™ flow cytometer (BD Biosciences) and analyzed using FlowJo analysis software (Treestar, Ashland, OR, USA).

    Recombinase Polymerase Amplification:

    Article Title: Intact CD100–CD72 Interaction Necessary for TCR-Induced T Cell Proliferation
    Article Snippet: .. Flow Cytometry Fresh or thawed PBMCs were stained for cell surface markers with fluorochrome-conjugated antibodies against the following proteins (dilutions indicated): CD3 1:200 (UCHT1, eBioscience), CD19 1:100 (HIB19, BD Biosciences), CD45 1:100 (HI30, BD Biosciences), CD4 1:100 (RPA-T4, eBioscience), CD8a 1:200 (RPA-T8, eBioscience), CD100 1:50 (A8, Biolegend), CD100 1:50 (unconjugated, 133/1C6, eBioscience), mouse IgM 1:100 (secondary ab for anti-CD100, eB121-15F9, eBioscience), mouse IgG 1:100 (secondary ab for other anti-CD100, Sigma-Aldrich), CD72 1:100 (3F3, Biolegend), and dead cell marker (DCM) 1:200 (Zombie NIR™ Fixable Viability Kit, Biolegend). .. Data were acquired on a FACSVerse™ flow cytometer (BD Biosciences) or on a LSRFortessa™ flow cytometer (BD Biosciences) and analyzed using FlowJo analysis software (Treestar, Ashland, OR, USA).

    In Vivo:

    Article Title: The BiP Cochaperone ERdj4 Is Required for B Cell Development and Function
    Article Snippet: .. B Cell function in vivo Basal immunoglobulins were measured in the serum of adult mice using the mouse IgM, IgG or IgA Ready-SET-Go ELISAs (Ebioscience) or the mouse IgE ELISA MAX™ Deluxe (Biolegend). .. To assess antigen-specific immunoglobulin production, mice were intraperitoneally injected with either 36 TNP-Ficoll (Biosearch Technologies, 50 µg) or 21 TNP-CGG (Biosearch Technologies, 100 µg) in Imject Alum adjuvant (Pierce, 1∶1 antigen to adjuvant ratio).

    Cytometry:

    Article Title: Intact CD100–CD72 Interaction Necessary for TCR-Induced T Cell Proliferation
    Article Snippet: .. Flow Cytometry Fresh or thawed PBMCs were stained for cell surface markers with fluorochrome-conjugated antibodies against the following proteins (dilutions indicated): CD3 1:200 (UCHT1, eBioscience), CD19 1:100 (HIB19, BD Biosciences), CD45 1:100 (HI30, BD Biosciences), CD4 1:100 (RPA-T4, eBioscience), CD8a 1:200 (RPA-T8, eBioscience), CD100 1:50 (A8, Biolegend), CD100 1:50 (unconjugated, 133/1C6, eBioscience), mouse IgM 1:100 (secondary ab for anti-CD100, eB121-15F9, eBioscience), mouse IgG 1:100 (secondary ab for other anti-CD100, Sigma-Aldrich), CD72 1:100 (3F3, Biolegend), and dead cell marker (DCM) 1:200 (Zombie NIR™ Fixable Viability Kit, Biolegend). .. Data were acquired on a FACSVerse™ flow cytometer (BD Biosciences) or on a LSRFortessa™ flow cytometer (BD Biosciences) and analyzed using FlowJo analysis software (Treestar, Ashland, OR, USA).

    Purification:

    Article Title: Expression analysis of surface molecules on human thymic dendritic cells with the 10th HLDA Workshop antibody panel
    Article Snippet: .. MAb recognizing human CD3 (OKT3, ATCC CRL-8001), CD11b (OKM1, ATCC CRL-8026), CD19 (BU12 ), HLA-DR (L234, ATCC HB-55) or mouse IgM (Bet-2) were purified from hybridoma supernatants and coupled to PE, Pacific Blue or Alexa Fluor 700 (Molecular Probes, Eugene, OR, USA) by standard procedures. .. For flow cytometry, analysis gates were set on live cells defined by scatter characteristics and exclusion of propidium iodide-positive events.

    Enzyme-linked Immunosorbent Assay:

    Article Title: The BiP Cochaperone ERdj4 Is Required for B Cell Development and Function
    Article Snippet: .. B Cell function in vivo Basal immunoglobulins were measured in the serum of adult mice using the mouse IgM, IgG or IgA Ready-SET-Go ELISAs (Ebioscience) or the mouse IgE ELISA MAX™ Deluxe (Biolegend). .. To assess antigen-specific immunoglobulin production, mice were intraperitoneally injected with either 36 TNP-Ficoll (Biosearch Technologies, 50 µg) or 21 TNP-CGG (Biosearch Technologies, 100 µg) in Imject Alum adjuvant (Pierce, 1∶1 antigen to adjuvant ratio).

    Incubation:

    Article Title: Mucosal immunoglobulins protect the olfactory organ of teleost fish against parasitic infection
    Article Snippet: .. Thereafter, the membranes were blocked with 8% skim milk and incubated with anti-trout IgT (rabbit pAb), anti-trout IgM (mouse monoclonal antibody (mAb)) or biotinylated anti-trout IgD (mouse mAb) antibodies followed by incubation with peroxidase-conjugated anti-rabbit, anti-mouse IgG (Invitrogen) or streptavidin (Invitrogen). .. Immunoreactivity was detected with an enhanced chemiluminescent reagent (Advansta) and scanned by GE Amersham Imager 600 Imaging System (GE Healthcare).

    other:

    Article Title: Multiplex Immunoassay Platforms Based on Shape-Coded Poly(ethylene glycol) Hydrogel Microparticles Incorporating Acrylic Acid
    Article Snippet: FITC-rabbit anti-mouse immunoglobulin G(FITC-anti-IgG), mouse IgG, FITC-rabbit anti-mouse immunoglobulin M(FITC-anti-IgM), and mouse IgM were purchased from ZYMED Laboratories (San Francisco, CA, USA).

    Confocal Microscopy:

    Article Title: Immunoglobulin M is Required for Protection Against Atherosclerosis in Low-Density Lipoprotein Receptor-Deficient Mice
    Article Snippet: .. For confocal microscopy, aortic root cryosections were double-immunostained for CD68 (Alexafluor 488-conjugated anti-CD68 Ab, Molecular Probes, Invitrogen, Paisley, UK) and IgM (biotin-conjugated mouse anti-mouse IgMb mAb and biotin-conjugated mouse anti-mouse IgMa , BD Pharmingen, Oxford, UK) secondarily labelled with Alexa 568-conjugated streptavidin (Molecular Probes), counterstained with TOPRO-3. .. Human LDL (density 1.019-1.063g/mL) was isolated from plasma of healthy donors after overnight fasting by differential density ultracentrifugation, and modified with either freshly synthesized malondialdehyde (MDA) or CuSO4 to generate MDA-LDL and copper-oxidised LDL (CuOxLDL) (see ).

    Marker:

    Article Title: Intact CD100–CD72 Interaction Necessary for TCR-Induced T Cell Proliferation
    Article Snippet: .. Flow Cytometry Fresh or thawed PBMCs were stained for cell surface markers with fluorochrome-conjugated antibodies against the following proteins (dilutions indicated): CD3 1:200 (UCHT1, eBioscience), CD19 1:100 (HIB19, BD Biosciences), CD45 1:100 (HI30, BD Biosciences), CD4 1:100 (RPA-T4, eBioscience), CD8a 1:200 (RPA-T8, eBioscience), CD100 1:50 (A8, Biolegend), CD100 1:50 (unconjugated, 133/1C6, eBioscience), mouse IgM 1:100 (secondary ab for anti-CD100, eB121-15F9, eBioscience), mouse IgG 1:100 (secondary ab for other anti-CD100, Sigma-Aldrich), CD72 1:100 (3F3, Biolegend), and dead cell marker (DCM) 1:200 (Zombie NIR™ Fixable Viability Kit, Biolegend). .. Data were acquired on a FACSVerse™ flow cytometer (BD Biosciences) or on a LSRFortessa™ flow cytometer (BD Biosciences) and analyzed using FlowJo analysis software (Treestar, Ashland, OR, USA).

    Staining:

    Article Title: Intact CD100–CD72 Interaction Necessary for TCR-Induced T Cell Proliferation
    Article Snippet: .. Flow Cytometry Fresh or thawed PBMCs were stained for cell surface markers with fluorochrome-conjugated antibodies against the following proteins (dilutions indicated): CD3 1:200 (UCHT1, eBioscience), CD19 1:100 (HIB19, BD Biosciences), CD45 1:100 (HI30, BD Biosciences), CD4 1:100 (RPA-T4, eBioscience), CD8a 1:200 (RPA-T8, eBioscience), CD100 1:50 (A8, Biolegend), CD100 1:50 (unconjugated, 133/1C6, eBioscience), mouse IgM 1:100 (secondary ab for anti-CD100, eB121-15F9, eBioscience), mouse IgG 1:100 (secondary ab for other anti-CD100, Sigma-Aldrich), CD72 1:100 (3F3, Biolegend), and dead cell marker (DCM) 1:200 (Zombie NIR™ Fixable Viability Kit, Biolegend). .. Data were acquired on a FACSVerse™ flow cytometer (BD Biosciences) or on a LSRFortessa™ flow cytometer (BD Biosciences) and analyzed using FlowJo analysis software (Treestar, Ashland, OR, USA).

    Article Title: Differential Impact of Chronic Hyperglycemia on Humoral Versus Cellular Primary Alloimmunity
    Article Snippet: .. Cells were then stained with secondary dye–conjugated anti-mouse IgM (Il/41) and anti-IgG antibody (11-4011-85; eBioscience). .. Data were graphed and statistically analyzed using Prism software.

    Mouse Assay:

    Article Title: The BiP Cochaperone ERdj4 Is Required for B Cell Development and Function
    Article Snippet: .. B Cell function in vivo Basal immunoglobulins were measured in the serum of adult mice using the mouse IgM, IgG or IgA Ready-SET-Go ELISAs (Ebioscience) or the mouse IgE ELISA MAX™ Deluxe (Biolegend). .. To assess antigen-specific immunoglobulin production, mice were intraperitoneally injected with either 36 TNP-Ficoll (Biosearch Technologies, 50 µg) or 21 TNP-CGG (Biosearch Technologies, 100 µg) in Imject Alum adjuvant (Pierce, 1∶1 antigen to adjuvant ratio).

    Cell Function Assay:

    Article Title: The BiP Cochaperone ERdj4 Is Required for B Cell Development and Function
    Article Snippet: .. B Cell function in vivo Basal immunoglobulins were measured in the serum of adult mice using the mouse IgM, IgG or IgA Ready-SET-Go ELISAs (Ebioscience) or the mouse IgE ELISA MAX™ Deluxe (Biolegend). .. To assess antigen-specific immunoglobulin production, mice were intraperitoneally injected with either 36 TNP-Ficoll (Biosearch Technologies, 50 µg) or 21 TNP-CGG (Biosearch Technologies, 100 µg) in Imject Alum adjuvant (Pierce, 1∶1 antigen to adjuvant ratio).

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  • 94
    Thermo Fisher mouse anti human igm secondary antibody
    Profiling seroconversion in COVID-19. Simoa serological assay results for <t>IgG,</t> <t>IgM</t> and IgA against the four viral targets (spike, S1 subunit, RBD and nucleocapsid) for pre-pandemic samples (light blue; n = 200), nasopharyngeal (NP) PCR-negative samples (dark blue; n = 100) and SARS-CoV-2-positive samples (black (immunocompetent; n = 141) or red (immunosuppressed; n = 31)). The SARS-CoV-2-positive samples were divided into four groups according to time since since the first positive nasopharyngeal RT–PCR test. Black lines indicate the median normalized AEB value of each population.
    Mouse Anti Human Igm Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher horseradish peroxidase conjugated goat anti mouse igm
    MHC-I and MHC-II are involved in antibody isotype switching. B2m KO, MHC-II KO, and WT C57BL/6 mice were vaccinated s.c. with 10 μg of PIV and challenged i.p. with 1 × 10 7 genomic copies of C. burnetii NMI 28 dpv. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. C. burnetii NMI-specific serum <t>IgM</t> (A to C) and <t>IgG</t> (D to F) were evaluated weekly following vaccination. Specific IgM (G) and IgG (H) were also evaluated 14 dpi. Each experimental group includes five mice, with error bars representing the standard deviations from the mean. **, P
    Horseradish Peroxidase Conjugated Goat Anti Mouse Igm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse igm secondary antibody
    The immunogenicity of M2FA-lysine-BSA in the absence of adjuvant and anti-M2FA antibody titers in intact mice. C57BL/6 mice were injected i.p. with M2FA-lysine-BSA or BSA in the absence of adjuvant. The antibody titers of <t>IgG</t> ( a ) and <t>IgM</t> ( b ) against M2FA-lysine were detected using MFA-6ACA-KLH-coated plates. The anti-M2AA antibody titers were clearly increased in M2FA-lysine-BSA-immunized mice compared to the controls (BSA-treated mice). Values are mean and SD. ( c ) Intact female C57BL/6 mice (n = 4 or 5 per group) with different ages showed significantly different anti-M2FA IgG titers. Values are mean and SD. (*p
    Goat Anti Mouse Igm Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Profiling seroconversion in COVID-19. Simoa serological assay results for IgG, IgM and IgA against the four viral targets (spike, S1 subunit, RBD and nucleocapsid) for pre-pandemic samples (light blue; n = 200), nasopharyngeal (NP) PCR-negative samples (dark blue; n = 100) and SARS-CoV-2-positive samples (black (immunocompetent; n = 141) or red (immunosuppressed; n = 31)). The SARS-CoV-2-positive samples were divided into four groups according to time since since the first positive nasopharyngeal RT–PCR test. Black lines indicate the median normalized AEB value of each population.

    Journal: Nature Biomedical Engineering

    Article Title: Ultrasensitive high-resolution profiling of early seroconversion in patients with COVID-19

    doi: 10.1038/s41551-020-00611-x

    Figure Lengend Snippet: Profiling seroconversion in COVID-19. Simoa serological assay results for IgG, IgM and IgA against the four viral targets (spike, S1 subunit, RBD and nucleocapsid) for pre-pandemic samples (light blue; n = 200), nasopharyngeal (NP) PCR-negative samples (dark blue; n = 100) and SARS-CoV-2-positive samples (black (immunocompetent; n = 141) or red (immunosuppressed; n = 31)). The SARS-CoV-2-positive samples were divided into four groups according to time since since the first positive nasopharyngeal RT–PCR test. Black lines indicate the median normalized AEB value of each population.

    Article Snippet: Anti-human immunoglobulin antibodies were diluted in Homebrew Detector/Sample Diluent to final concentrations of 7.73 ng ml−1 IgG (Bethyl Labratories; A80-148B), 216 ng ml−1 IgM (Thermo Fisher Scientific; MII0401) and 150 ng ml−1 IgA (Abcam; ab214003).

    Techniques: Serologic Assay, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

    Classification of COVID-19 using Simoa serological assays. a , Left: ROC curves for all positive cases ( n = 141) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green), the early-stage model (blue) or the late-stage model (red). Middle: ROC curves for late-stage cases (more than 1 week after positive nasopharyngeal PCR test; n = 50) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green) or the late-stage model (red). Right: ROC curves for early-stage cases (first week after positive nasopharyngeal PCR test; n = 91) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green) or the early-stage model (blue). AUC values and 95% confidence intervals (CIs) are shown for each graph. b , Simoa serological assay results for IgG, IgM and IgA against the four viral targets (spike, S1 subunit, RBD and nucleocapsid) for pre-pandemic samples (light blue; n = 232) and nasopharyngeal RT–PCR-positive samples (grey; n = 68). Statistical significance was determined using Mann–Whitney U -tests. All U -tests were two tailed and did not correct for multiple comparisons. Black lines indicate the median normalized AEB value of each population.

    Journal: Nature Biomedical Engineering

    Article Title: Ultrasensitive high-resolution profiling of early seroconversion in patients with COVID-19

    doi: 10.1038/s41551-020-00611-x

    Figure Lengend Snippet: Classification of COVID-19 using Simoa serological assays. a , Left: ROC curves for all positive cases ( n = 141) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green), the early-stage model (blue) or the late-stage model (red). Middle: ROC curves for late-stage cases (more than 1 week after positive nasopharyngeal PCR test; n = 50) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green) or the late-stage model (red). Right: ROC curves for early-stage cases (first week after positive nasopharyngeal PCR test; n = 91) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green) or the early-stage model (blue). AUC values and 95% confidence intervals (CIs) are shown for each graph. b , Simoa serological assay results for IgG, IgM and IgA against the four viral targets (spike, S1 subunit, RBD and nucleocapsid) for pre-pandemic samples (light blue; n = 232) and nasopharyngeal RT–PCR-positive samples (grey; n = 68). Statistical significance was determined using Mann–Whitney U -tests. All U -tests were two tailed and did not correct for multiple comparisons. Black lines indicate the median normalized AEB value of each population.

    Article Snippet: Anti-human immunoglobulin antibodies were diluted in Homebrew Detector/Sample Diluent to final concentrations of 7.73 ng ml−1 IgG (Bethyl Labratories; A80-148B), 216 ng ml−1 IgM (Thermo Fisher Scientific; MII0401) and 150 ng ml−1 IgA (Abcam; ab214003).

    Techniques: Marker, Polymerase Chain Reaction, Serologic Assay, Reverse Transcription Polymerase Chain Reaction, MANN-WHITNEY, Two Tailed Test

    Schematic of the Simoa serological assay. Plasma is incubated with four types of dye-encoded beads that are each coupled to one of four viral targets (spike, S1, RBD and nucleocapsid). IgG, IgA and IgM antibodies specific to the SARS-CoV-2 targets bind to the viral antigen-coated beads. After washing, beads are introduced to biotinylated anti-human immunoglobulin antibodies to label either IgG, IgM or IgA in the different reactions. After additional washes, the enzyme SβG is introduced. The beads are washed, resuspended in fluorogenic RGP and loaded into a 216,000-microwell array for multicolour imaging.

    Journal: Nature Biomedical Engineering

    Article Title: Ultrasensitive high-resolution profiling of early seroconversion in patients with COVID-19

    doi: 10.1038/s41551-020-00611-x

    Figure Lengend Snippet: Schematic of the Simoa serological assay. Plasma is incubated with four types of dye-encoded beads that are each coupled to one of four viral targets (spike, S1, RBD and nucleocapsid). IgG, IgA and IgM antibodies specific to the SARS-CoV-2 targets bind to the viral antigen-coated beads. After washing, beads are introduced to biotinylated anti-human immunoglobulin antibodies to label either IgG, IgM or IgA in the different reactions. After additional washes, the enzyme SβG is introduced. The beads are washed, resuspended in fluorogenic RGP and loaded into a 216,000-microwell array for multicolour imaging.

    Article Snippet: Anti-human immunoglobulin antibodies were diluted in Homebrew Detector/Sample Diluent to final concentrations of 7.73 ng ml−1 IgG (Bethyl Labratories; A80-148B), 216 ng ml−1 IgM (Thermo Fisher Scientific; MII0401) and 150 ng ml−1 IgA (Abcam; ab214003).

    Techniques: Serologic Assay, Incubation, Imaging

    MHC-I and MHC-II are involved in antibody isotype switching. B2m KO, MHC-II KO, and WT C57BL/6 mice were vaccinated s.c. with 10 μg of PIV and challenged i.p. with 1 × 10 7 genomic copies of C. burnetii NMI 28 dpv. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. C. burnetii NMI-specific serum IgM (A to C) and IgG (D to F) were evaluated weekly following vaccination. Specific IgM (G) and IgG (H) were also evaluated 14 dpi. Each experimental group includes five mice, with error bars representing the standard deviations from the mean. **, P

    Journal: Infection and Immunity

    Article Title: Major Histocompatibility Complex Class II-Restricted, CD4+ T Cell-Dependent and -Independent Mechanisms Are Required for Vaccine-Induced Protective Immunity against Coxiella burnetii

    doi: 10.1128/IAI.00824-19

    Figure Lengend Snippet: MHC-I and MHC-II are involved in antibody isotype switching. B2m KO, MHC-II KO, and WT C57BL/6 mice were vaccinated s.c. with 10 μg of PIV and challenged i.p. with 1 × 10 7 genomic copies of C. burnetii NMI 28 dpv. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. C. burnetii NMI-specific serum IgM (A to C) and IgG (D to F) were evaluated weekly following vaccination. Specific IgM (G) and IgG (H) were also evaluated 14 dpi. Each experimental group includes five mice, with error bars representing the standard deviations from the mean. **, P

    Article Snippet: Plates were washed with PBS-T buffer and then incubated with 100 μl of diluted horseradish peroxidase-conjugated goat anti-mouse IgM or IgG (1:4,000 to 1:8,000) at room temperature for 1 h. The plates were again washed with PBS-T, followed by the addition of 100 μl of TMB substrate (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Mouse Assay

    CD4 + T cells are important for antibody isotype switching. WT, CD4 KO, and MHC-II KO mice were vaccinated and challenged as previously described. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. C. burnetii NMI-specific serum IgM and IgG were evaluated weekly until 28 dpv (A) and 14 dpi (B). Each experimental group includes five mice, with error bars representing the standard deviations from the mean. *, P

    Journal: Infection and Immunity

    Article Title: Major Histocompatibility Complex Class II-Restricted, CD4+ T Cell-Dependent and -Independent Mechanisms Are Required for Vaccine-Induced Protective Immunity against Coxiella burnetii

    doi: 10.1128/IAI.00824-19

    Figure Lengend Snippet: CD4 + T cells are important for antibody isotype switching. WT, CD4 KO, and MHC-II KO mice were vaccinated and challenged as previously described. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. C. burnetii NMI-specific serum IgM and IgG were evaluated weekly until 28 dpv (A) and 14 dpi (B). Each experimental group includes five mice, with error bars representing the standard deviations from the mean. *, P

    Article Snippet: Plates were washed with PBS-T buffer and then incubated with 100 μl of diluted horseradish peroxidase-conjugated goat anti-mouse IgM or IgG (1:4,000 to 1:8,000) at room temperature for 1 h. The plates were again washed with PBS-T, followed by the addition of 100 μl of TMB substrate (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Mouse Assay

    The immunogenicity of M2FA-lysine-BSA in the absence of adjuvant and anti-M2FA antibody titers in intact mice. C57BL/6 mice were injected i.p. with M2FA-lysine-BSA or BSA in the absence of adjuvant. The antibody titers of IgG ( a ) and IgM ( b ) against M2FA-lysine were detected using MFA-6ACA-KLH-coated plates. The anti-M2AA antibody titers were clearly increased in M2FA-lysine-BSA-immunized mice compared to the controls (BSA-treated mice). Values are mean and SD. ( c ) Intact female C57BL/6 mice (n = 4 or 5 per group) with different ages showed significantly different anti-M2FA IgG titers. Values are mean and SD. (*p

    Journal: Scientific Reports

    Article Title: Evidence that endogenous formaldehyde produces immunogenic and atherogenic adduct epitopes

    doi: 10.1038/s41598-017-11289-8

    Figure Lengend Snippet: The immunogenicity of M2FA-lysine-BSA in the absence of adjuvant and anti-M2FA antibody titers in intact mice. C57BL/6 mice were injected i.p. with M2FA-lysine-BSA or BSA in the absence of adjuvant. The antibody titers of IgG ( a ) and IgM ( b ) against M2FA-lysine were detected using MFA-6ACA-KLH-coated plates. The anti-M2AA antibody titers were clearly increased in M2FA-lysine-BSA-immunized mice compared to the controls (BSA-treated mice). Values are mean and SD. ( c ) Intact female C57BL/6 mice (n = 4 or 5 per group) with different ages showed significantly different anti-M2FA IgG titers. Values are mean and SD. (*p

    Article Snippet: Malondialdehyde bis(dimethyl) acetal, Dynabeads M-270 Amine, CarboxyLink Kit, the Imject EDC mcKLH Spin Kit, goat anti-rabbit IgG (H + L) antibody with HRP, goat anti-human IgG/IgM secondary antibody, and goat anti-mouse IgM secondary antibody were obtained from Thermo Scientific (Rockford, IL).

    Techniques: Mouse Assay, Injection

    Serum anti-M2FA IgG and IgM antibody levels in wild-type and ApoE −/− mice and immunohistochemical detection of M2FA-epitopes in heart valve of ApoE −/− mice. The anti-M2FA IgG ( a ) and IgM ( b ) antibody levels showed significant differences between wild-type and ApoE −/− mice with the M2FA-6ACA-BSA ELISAs (**p

    Journal: Scientific Reports

    Article Title: Evidence that endogenous formaldehyde produces immunogenic and atherogenic adduct epitopes

    doi: 10.1038/s41598-017-11289-8

    Figure Lengend Snippet: Serum anti-M2FA IgG and IgM antibody levels in wild-type and ApoE −/− mice and immunohistochemical detection of M2FA-epitopes in heart valve of ApoE −/− mice. The anti-M2FA IgG ( a ) and IgM ( b ) antibody levels showed significant differences between wild-type and ApoE −/− mice with the M2FA-6ACA-BSA ELISAs (**p

    Article Snippet: Malondialdehyde bis(dimethyl) acetal, Dynabeads M-270 Amine, CarboxyLink Kit, the Imject EDC mcKLH Spin Kit, goat anti-rabbit IgG (H + L) antibody with HRP, goat anti-human IgG/IgM secondary antibody, and goat anti-mouse IgM secondary antibody were obtained from Thermo Scientific (Rockford, IL).

    Techniques: Mouse Assay, Immunohistochemistry