monoclonal anti human immunoglobulin m igm  (Thermo Fisher)


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    Name:
    Human IgM Monoclonal Antibody SA DA4
    Description:
    Human IgM Monoclonal Antibody for IHC Flow
    Catalog Number:
    11-9998-41
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
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    Structured Review

    Thermo Fisher monoclonal anti human immunoglobulin m igm
    Western blot of P. knowlesi pooled sera probed with P. vivax pooled sera. Unfractionated, pooled serum samples from patient is subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human <t>IgM-HRP</t> (as secondary antibody).
    Human IgM Monoclonal Antibody for IHC Flow
    https://www.bioz.com/result/monoclonal anti human immunoglobulin m igm/product/Thermo Fisher
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti human immunoglobulin m igm - by Bioz Stars, 2021-06
    92/100 stars

    Images

    1) Product Images from "Identification of circulating biomarkers in sera of Plasmodium knowlesi-infected malaria patients – comparison against Plasmodium vivax infection"

    Article Title: Identification of circulating biomarkers in sera of Plasmodium knowlesi-infected malaria patients – comparison against Plasmodium vivax infection

    Journal: BMC Infectious Diseases

    doi: 10.1186/s12879-015-0786-2

    Western blot of P. knowlesi pooled sera probed with P. vivax pooled sera. Unfractionated, pooled serum samples from patient is subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).
    Figure Legend Snippet: Western blot of P. knowlesi pooled sera probed with P. vivax pooled sera. Unfractionated, pooled serum samples from patient is subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).

    Techniques Used: Western Blot

    Western blot results following 2-DE. (a) normal pooled sera probed with normal pooled sera, (b) normal pooled sera probed with P. knowlesi pooled sera, (c) P. knowlesi pooled sera probed with normal pooled sera, (d) P. knowlesi pooled sera probed with P. knowlesi pooled sera, Unfractionated, pooled serum samples from patients and normal controls were subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).
    Figure Legend Snippet: Western blot results following 2-DE. (a) normal pooled sera probed with normal pooled sera, (b) normal pooled sera probed with P. knowlesi pooled sera, (c) P. knowlesi pooled sera probed with normal pooled sera, (d) P. knowlesi pooled sera probed with P. knowlesi pooled sera, Unfractionated, pooled serum samples from patients and normal controls were subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).

    Techniques Used: Western Blot

    Related Articles

    Incubation:

    Article Title: Efficient Acquisition of Fully Human Antibody Genes against Self-Proteins by Sorting Single B Cells Stimulated with Vaccines Based on Nitrated T Helper Cell Epitopes
    Article Snippet: After washing 5 times with PBST (PBS containing 0.05% Tween 20), the plates were blocked with PBS containing 6% BSA (BioFroxx, Germany) at 37°C for 2 h. The serum was diluted 400 or 800 times with PBS containing 2% BSA. .. Then, the plates were incubated with diluted serum at 37°C for 2 h. After washing 5 times with PBST, the plates were incubated with HRP-conjugated goat antibodies (1 : 10000 v /v ) against human IgM or IgG (Thermo Fisher Scientific, USA) at 37°C for 1 h. Finally, the TMB substrate chromogenic solution (Solarbio, China) was added and incubated at 37°C for 15 min. ..

    Article Title: Identification of circulating biomarkers in sera of Plasmodium knowlesi-infected malaria patients – comparison against Plasmodium vivax infection
    Article Snippet: The membranes were subsequently incubated overnight (4°C) with the indicated primary antibodies, which corresponded to P. knowlesi infection, P. vivax infection, or normal healthy controls (all diluted at 1:50). .. After another washing step, the membranes were incubated with monoclonal anti-human Immunoglobulin M (IgM) conjugated to horseradish peroxidase (HRP) (Invitrogen, USA) at a dilution of 1:6,000 for 1 h at room temperature. .. The resulting immunocomplexes were visualized using chemiluminescent blotting reagent (Pierce, USA) and X-ray film (18 × 24 cm; Kodak).

    Indirect ELISA:

    Article Title: Amyloid-? Oligomer Specificity Mediated by the IgM Isotype - Implications for a Specific Protective Mechanism Exerted by Endogenous Auto-Antibodies
    Article Snippet: Blocking solution and antibody-dilutions were made with 5% Non-fat dry milk in PBST and all washes were performed with PBS containing 0.1% Tween-20 (PBST). .. Indirect ELISA using anti-human IgM The anti-human IgM antibody (Thermo scientific, Rockford, USA), diluted to 10 µg/ml in PBS, was adsorbed to Nunc-Immuno MaxiSorp plates (Nunc, Roskilde, Denmark). .. Human plasma from eight anonymous healthy donors was diluted 4X in PBS, added to the wells, and incubated for 1 hour at RT.

    Labeling:

    Article Title: Humoral anti-KLH responses in cancer patients treated with dendritic cell-based immunotherapy are dictated by different vaccination parameters
    Article Snippet: To prepare suitable standard curves for the anti-KLH ELISA’s, the work standard was serially diluted to establish a 7-point standard curve for each Ab isotype (Supplementary Figure 1, available online). .. After washing, specific Abs against human IgM or IgA or total IgG or IgG1 or IgG2 or IgG3 or IgG4 labeled with horseradish peroxidase (Invitrogen, San Diego, CA, USA) at 1:500 were added to the wells as 100 μL aliquots. ..

    Concentration Assay:

    Article Title: Mannose-Containing Oligosaccharides of Non-Specific Human Secretory Immunoglobulin A Mediate Inhibition of Vibrio cholerae Biofilm Formation
    Article Snippet: Briefly, V. cholerae 0139 (MO10) and the vspR mutant were grown overnight at 37° C with shaking in LB containing 50 µg/ml streptomycin. .. The bacterial cultures were diluted in 2X LB to a concentration of approximately 2×106 CFU/ml and 50 µL was inoculated into sterile 96-well polystyrene microtiter plates along with 50 µL of PBS, or bovine serum albumin (BSA) (Fisher Scientific, Fairlawn, NJ) or serum immunoglobulin A (IgA, Pierce, Rockford, IL) or secretory IgA (SIgA) (Sigma Aldrich) or human serum IgG (Pierce) or human serum IgM (Pierce) at a final concentration of 200 µg/well in PBS. .. In another experiment, escalating doses of free mannose (Difco, Detroit, MI) (serially doubling from 0.0015% to 0.03%) were added to cultures of MO10.

    Purification:

    Article Title: IgM, IgG, and IgA Antibody Responses to Influenza A(H1N1)pdm09 Hemagglutinin in Infected Persons during the First Wave of the 2009 Pandemic in the United States
    Article Snippet: Purified, trimeric HA was captured via the C-terminal 6-histidine tag on HisGrab nickel-coated plates (Thermo Fisher Scientific, IL) at 200 ng/well. .. Standard curves were prepared using purified human IgM, IgG, and IgA standards (Thermo Fisher Scientific, IL). ..

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  • 92
    Thermo Fisher monoclonal anti human immunoglobulin m igm
    Western blot of P. knowlesi pooled sera probed with P. vivax pooled sera. Unfractionated, pooled serum samples from patient is subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human <t>IgM-HRP</t> (as secondary antibody).
    Monoclonal Anti Human Immunoglobulin M Igm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti human immunoglobulin m igm/product/Thermo Fisher
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti human immunoglobulin m igm - by Bioz Stars, 2021-06
    92/100 stars
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    94
    Thermo Fisher mab mouse anti trout igm
    Ig-specific responses in the pharynx from infected and survivor fish. ( A and B ) Four different microscope images of slides showing immunofluorescence staining of I. multifiliis parasites in pharyngeal paraffinic sections from trout infected with I. multifiliis after 28 d ( n = 6). (A) From left to right: I. multifiliis (magenta), <t>IgM</t> (red), and <t>IgT</t> (green), with nuclei stained with DAPI (blue). (B) From left to right: I. multifiliis ). Scale bar, 20 μm. ( C and D ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in pharyngeal mucus (dilution 1:2) from 28 d–infected (C) and survivor (D) fish. ( E and F ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in serum (dilution 1:10) from 28 d–infected (E) and survivor (F) fish. ( G and H ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of pharyngeal mucus from infected (G) and survivor (H) fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control uninfected fish ( n = 12). ( I and J ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of serum from 28 d–infected (I) and survivor (J) fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control uninfected fish ( n = 12). Data are representative of at least three independent experiments (mean ± SEM). * p
    Mab Mouse Anti Trout Igm, 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
    https://www.bioz.com/result/mab mouse anti trout igm/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    90
    Thermo Fisher peroxidase conjugated anti mouse igm
    OmpC and OmpF from S . Typhi promote antibody responses to ovalbumin (OVA) . Four BALB/c mice per group were immunized i.p. with 100 μg of OVA, OVA + 10 μg of porins, OVA + 10 μg of PorK, OVA + 5 μg of LPS, or OVA + 100 μg of alum. (A) Total <t>IgG</t> antibody responses were measured at the indicated time points by ELISA (B) high-avidity IgG and (C) IgG1, IgG2a, and IgG2b were measured at day 30 post-immunization. (D) BALB/c mice were injected i.p. with a depleting anti-CD4 mAb (GK1.5) 3 days prior to immunization with OVA or OVA + porins. GK1.5 mAb was given every 3 days thereafter until day 20, and the serum IgG and <t>IgM</t> responses were measured by ELISA. Mean + SEM are shown. These data are representative of three independent experiments. Statistical analysis was performed using one-way ANOVA with Bonferroni test post hoc . For panel (A) , Student’s t -test was performed comparing only OVA versus OVA + porins groups. Statistical differences are depicted as * P
    Peroxidase Conjugated Anti Mouse Igm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase conjugated anti mouse igm/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
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    93
    Thermo Fisher anti cd40 monoclonal antibody
    The production of IL-10 by PBMC or CD8-depleted cells (– CD8) from individuals with Graves’ disease or Hashimoto’s thyroiditis, and normal controls. The PBMC or CD8-depleted cells were incubated in the presence or absence of <t>anti-CD40</t> MoAb and IL-4. * P
    Anti Cd40 Monoclonal Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Western blot of P. knowlesi pooled sera probed with P. vivax pooled sera. Unfractionated, pooled serum samples from patient is subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).

    Journal: BMC Infectious Diseases

    Article Title: Identification of circulating biomarkers in sera of Plasmodium knowlesi-infected malaria patients – comparison against Plasmodium vivax infection

    doi: 10.1186/s12879-015-0786-2

    Figure Lengend Snippet: Western blot of P. knowlesi pooled sera probed with P. vivax pooled sera. Unfractionated, pooled serum samples from patient is subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).

    Article Snippet: After another washing step, the membranes were incubated with monoclonal anti-human Immunoglobulin M (IgM) conjugated to horseradish peroxidase (HRP) (Invitrogen, USA) at a dilution of 1:6,000 for 1 h at room temperature.

    Techniques: Western Blot

    Western blot results following 2-DE. (a) normal pooled sera probed with normal pooled sera, (b) normal pooled sera probed with P. knowlesi pooled sera, (c) P. knowlesi pooled sera probed with normal pooled sera, (d) P. knowlesi pooled sera probed with P. knowlesi pooled sera, Unfractionated, pooled serum samples from patients and normal controls were subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).

    Journal: BMC Infectious Diseases

    Article Title: Identification of circulating biomarkers in sera of Plasmodium knowlesi-infected malaria patients – comparison against Plasmodium vivax infection

    doi: 10.1186/s12879-015-0786-2

    Figure Lengend Snippet: Western blot results following 2-DE. (a) normal pooled sera probed with normal pooled sera, (b) normal pooled sera probed with P. knowlesi pooled sera, (c) P. knowlesi pooled sera probed with normal pooled sera, (d) P. knowlesi pooled sera probed with P. knowlesi pooled sera, Unfractionated, pooled serum samples from patients and normal controls were subjected to 2-DE, transferred onto nitrocellulose membranes, and probed with pooled sera (as primary antibody) followed by monoclonal anti-human IgM-HRP (as secondary antibody).

    Article Snippet: After another washing step, the membranes were incubated with monoclonal anti-human Immunoglobulin M (IgM) conjugated to horseradish peroxidase (HRP) (Invitrogen, USA) at a dilution of 1:6,000 for 1 h at room temperature.

    Techniques: Western Blot

    Ig-specific responses in the pharynx from infected and survivor fish. ( A and B ) Four different microscope images of slides showing immunofluorescence staining of I. multifiliis parasites in pharyngeal paraffinic sections from trout infected with I. multifiliis after 28 d ( n = 6). (A) From left to right: I. multifiliis (magenta), IgM (red), and IgT (green), with nuclei stained with DAPI (blue). (B) From left to right: I. multifiliis ). Scale bar, 20 μm. ( C and D ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in pharyngeal mucus (dilution 1:2) from 28 d–infected (C) and survivor (D) fish. ( E and F ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in serum (dilution 1:10) from 28 d–infected (E) and survivor (F) fish. ( G and H ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of pharyngeal mucus from infected (G) and survivor (H) fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control uninfected fish ( n = 12). ( I and J ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of serum from 28 d–infected (I) and survivor (J) fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control uninfected fish ( n = 12). Data are representative of at least three independent experiments (mean ± SEM). * p

    Journal: The Journal of Immunology Author Choice

    Article Title: Pharyngeal Immunity in Early Vertebrates Provides Functional and Evolutionary Insight into Mucosal Homeostasis

    doi: 10.4049/jimmunol.1900863

    Figure Lengend Snippet: Ig-specific responses in the pharynx from infected and survivor fish. ( A and B ) Four different microscope images of slides showing immunofluorescence staining of I. multifiliis parasites in pharyngeal paraffinic sections from trout infected with I. multifiliis after 28 d ( n = 6). (A) From left to right: I. multifiliis (magenta), IgM (red), and IgT (green), with nuclei stained with DAPI (blue). (B) From left to right: I. multifiliis ). Scale bar, 20 μm. ( C and D ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in pharyngeal mucus (dilution 1:2) from 28 d–infected (C) and survivor (D) fish. ( E and F ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in serum (dilution 1:10) from 28 d–infected (E) and survivor (F) fish. ( G and H ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of pharyngeal mucus from infected (G) and survivor (H) fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control uninfected fish ( n = 12). ( I and J ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of serum from 28 d–infected (I) and survivor (J) fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control uninfected fish ( n = 12). Data are representative of at least three independent experiments (mean ± SEM). * p

    Article Snippet: After 24 h, leukocytes from pharyngeal tissue or head kidney were obtained as described above, and cells were incubated with mAb mouse anti-trout IgM or anti-trout IgT (1 μg/ml each) on ice for 1 h. After washing three times, Alexa Fluor 488 goat anti-mouse IgG1 or Alexa Fluor 488 goat anti-mouse IgG2b (3 μg/ml each; Invitrogen) were used as secondary Ab to detect IgM+ or IgT+ B cells, respectively.

    Techniques: Infection, Fluorescence In Situ Hybridization, Microscopy, Immunofluorescence, Staining, Western Blot, Binding Assay

    A majority of trout pharyngeal bacteria are predominantly coated with IgT. ( A ) Representative flow cytometry histograms showing the staining of pharyngeal bacteria with IgT, IgM, and IgD. Bacteria were stained with isotype controls (shaded histograms), anti-trout IgT (green line), anti-trout IgM (red line), or anti-trout IgD (magenta line) mAbs, respectively. ( B ) Percentages of pharyngeal bacteria coated with IgT, IgM, or IgD ( n = 25). The median percentage is showed by a red line. ( C ) Immunoblot analysis of IgT, IgM, or IgD coating on pharyngeal bacteria. Lane 1, 0.1 μg of purified IgT, IgM, or IgD; lanes 2–7, pharyngeal bacteria ( n = 6). ( D ) Percentages of total pharyngeal mucus IgT, IgM, or IgD coating pharyngeal bacteria ( n = 12). The median is shown by a red line. The statistical differences in (B) and (D) were evaluated by one-way ANOVA with Bonferroni correction. Data are representative of at least three independent experiments. ( E ). Scale bar, 5 μm.

    Journal: The Journal of Immunology Author Choice

    Article Title: Pharyngeal Immunity in Early Vertebrates Provides Functional and Evolutionary Insight into Mucosal Homeostasis

    doi: 10.4049/jimmunol.1900863

    Figure Lengend Snippet: A majority of trout pharyngeal bacteria are predominantly coated with IgT. ( A ) Representative flow cytometry histograms showing the staining of pharyngeal bacteria with IgT, IgM, and IgD. Bacteria were stained with isotype controls (shaded histograms), anti-trout IgT (green line), anti-trout IgM (red line), or anti-trout IgD (magenta line) mAbs, respectively. ( B ) Percentages of pharyngeal bacteria coated with IgT, IgM, or IgD ( n = 25). The median percentage is showed by a red line. ( C ) Immunoblot analysis of IgT, IgM, or IgD coating on pharyngeal bacteria. Lane 1, 0.1 μg of purified IgT, IgM, or IgD; lanes 2–7, pharyngeal bacteria ( n = 6). ( D ) Percentages of total pharyngeal mucus IgT, IgM, or IgD coating pharyngeal bacteria ( n = 12). The median is shown by a red line. The statistical differences in (B) and (D) were evaluated by one-way ANOVA with Bonferroni correction. Data are representative of at least three independent experiments. ( E ). Scale bar, 5 μm.

    Article Snippet: After 24 h, leukocytes from pharyngeal tissue or head kidney were obtained as described above, and cells were incubated with mAb mouse anti-trout IgM or anti-trout IgT (1 μg/ml each) on ice for 1 h. After washing three times, Alexa Fluor 488 goat anti-mouse IgG1 or Alexa Fluor 488 goat anti-mouse IgG2b (3 μg/ml each; Invitrogen) were used as secondary Ab to detect IgM+ or IgT+ B cells, respectively.

    Techniques: Flow Cytometry, Cytometry, Staining, Purification

    Proliferative responses of IgT + and IgM + B cells in the pharynx of survived fish. ( A and B ) Immunofluorescence analysis of EdU incorporation by IgT + or IgM + B cells in the pharynx of control (A) and survivor fish (B). Pharynx paraffinic sections were stained for EdU (magenta), trout IgT (green), trout IgM (red), and nuclei (blue) detection ( n = 9). White rectangle represents partial enlargement area. White arrowheads point to cells double stained for EdU and IgT. Data are representative of at least three independent experiments. Scale bar, 20 μm. ( C ) Percentage of EdU + cells from the total pharynx IgT + or IgM + B cell populations in control or survivor fish counted from (A) and (B). ( D and E ) Representative flow cytometry dot plot showing proliferation of IgT + (D) and IgM + (E) B cells in pharynx leukocytes of control and survivor fish. The percentage of lymphocytes representing proliferative B cells (EdU + ) is shown in each dot plot. ( F ) Percentage of EdU + cells from the total pharynx IgT + or IgM + B cell populations in control and survivor fish ( n = 9). ( G and H ) Representative flow cytometry dot plot showing proliferation of IgT + (G) and IgM + (H) B cells in head kidney leukocytes of control and survivor fish. The percentage of lymphocytes representing proliferative B cells (EdU + ) is shown in each dot plot. ( I ) Percentages of EdU + cells from the total head kidney IgT + or IgM + B cell populations in control and survivor fish ( n = 9). Data in (C), (F), and (I) are representative of at least three independent experiments (mean ± SEM). ** p

    Journal: The Journal of Immunology Author Choice

    Article Title: Pharyngeal Immunity in Early Vertebrates Provides Functional and Evolutionary Insight into Mucosal Homeostasis

    doi: 10.4049/jimmunol.1900863

    Figure Lengend Snippet: Proliferative responses of IgT + and IgM + B cells in the pharynx of survived fish. ( A and B ) Immunofluorescence analysis of EdU incorporation by IgT + or IgM + B cells in the pharynx of control (A) and survivor fish (B). Pharynx paraffinic sections were stained for EdU (magenta), trout IgT (green), trout IgM (red), and nuclei (blue) detection ( n = 9). White rectangle represents partial enlargement area. White arrowheads point to cells double stained for EdU and IgT. Data are representative of at least three independent experiments. Scale bar, 20 μm. ( C ) Percentage of EdU + cells from the total pharynx IgT + or IgM + B cell populations in control or survivor fish counted from (A) and (B). ( D and E ) Representative flow cytometry dot plot showing proliferation of IgT + (D) and IgM + (E) B cells in pharynx leukocytes of control and survivor fish. The percentage of lymphocytes representing proliferative B cells (EdU + ) is shown in each dot plot. ( F ) Percentage of EdU + cells from the total pharynx IgT + or IgM + B cell populations in control and survivor fish ( n = 9). ( G and H ) Representative flow cytometry dot plot showing proliferation of IgT + (G) and IgM + (H) B cells in head kidney leukocytes of control and survivor fish. The percentage of lymphocytes representing proliferative B cells (EdU + ) is shown in each dot plot. ( I ) Percentages of EdU + cells from the total head kidney IgT + or IgM + B cell populations in control and survivor fish ( n = 9). Data in (C), (F), and (I) are representative of at least three independent experiments (mean ± SEM). ** p

    Article Snippet: After 24 h, leukocytes from pharyngeal tissue or head kidney were obtained as described above, and cells were incubated with mAb mouse anti-trout IgM or anti-trout IgT (1 μg/ml each) on ice for 1 h. After washing three times, Alexa Fluor 488 goat anti-mouse IgG1 or Alexa Fluor 488 goat anti-mouse IgG2b (3 μg/ml each; Invitrogen) were used as secondary Ab to detect IgM+ or IgT+ B cells, respectively.

    Techniques: Fluorescence In Situ Hybridization, Immunofluorescence, Staining, Flow Cytometry, Cytometry

    Accumulation of IgT + B cells in the pharynx of trout infected with I. multifiliis . ( A – C ) Representative differential interference contrast (DIC) images of immunofluorescence staining on paraffinic sections of pharynx from uninfected control fish (A), 28 d–infected fish (B), and survivor fish (C). IgT + and IgM + ). ( D ) Enlarged images of the areas outlined in (C) are showing some IgT + B cells possibly secreting IgT in pharyngeal epithelium (white arrowhead). Data are representative of at least three independent experiments ( n = 9 per group). Scale bar, 20 μm. ( E ) Numbers of B cells in control, 28 d–infected, and survivor fish counted from (A)–(C). ( F and G ) Concentration of IgT, IgM, and IgD in pharyngeal mucus (F) and serum (G) from uninfected control fish, 28 d–infected fish, and survivor fish ( n = 12). Data in (E)–(G) are representative of at least three independent experiments (mean ± SEM). * p

    Journal: The Journal of Immunology Author Choice

    Article Title: Pharyngeal Immunity in Early Vertebrates Provides Functional and Evolutionary Insight into Mucosal Homeostasis

    doi: 10.4049/jimmunol.1900863

    Figure Lengend Snippet: Accumulation of IgT + B cells in the pharynx of trout infected with I. multifiliis . ( A – C ) Representative differential interference contrast (DIC) images of immunofluorescence staining on paraffinic sections of pharynx from uninfected control fish (A), 28 d–infected fish (B), and survivor fish (C). IgT + and IgM + ). ( D ) Enlarged images of the areas outlined in (C) are showing some IgT + B cells possibly secreting IgT in pharyngeal epithelium (white arrowhead). Data are representative of at least three independent experiments ( n = 9 per group). Scale bar, 20 μm. ( E ) Numbers of B cells in control, 28 d–infected, and survivor fish counted from (A)–(C). ( F and G ) Concentration of IgT, IgM, and IgD in pharyngeal mucus (F) and serum (G) from uninfected control fish, 28 d–infected fish, and survivor fish ( n = 12). Data in (E)–(G) are representative of at least three independent experiments (mean ± SEM). * p

    Article Snippet: After 24 h, leukocytes from pharyngeal tissue or head kidney were obtained as described above, and cells were incubated with mAb mouse anti-trout IgM or anti-trout IgT (1 μg/ml each) on ice for 1 h. After washing three times, Alexa Fluor 488 goat anti-mouse IgG1 or Alexa Fluor 488 goat anti-mouse IgG2b (3 μg/ml each; Invitrogen) were used as secondary Ab to detect IgM+ or IgT+ B cells, respectively.

    Techniques: Infection, Immunofluorescence, Staining, Fluorescence In Situ Hybridization, Concentration Assay

    General organization of teleost PM lymphoid tissue. ( A ) H E stain of the pharynx of a control adult rainbow trout ( O. mykiss ). Black arrowhead indicates taste bud. ( B ) H E (upper) and Alcian blue (lower) stain of the pharynx from five different families of control adult teleost fish. From left to right: grass carp ( C. idellus ), southern catfish ( S. meridionalis ), mandarin fish ( S. chuatsi ), snakehead ( O. argus Cantor), and rainbow trout ( O. mykiss ). Scale bar, 50 μm. Black and red arrowheads indicate lymphocytes and mucus cells, respectively. ( C ) Heat map illustrates results from qPCR of mRNAs for selected immune markers in trout skin, gut, gill, pharynx, spleen, and head kidney ( n = 6). Data are expressed as mean cycle threshold (Ct) values ± SEM. ( D ) Flow cytometry analysis of pharynx (left) and head kidney (right) leukocytes stained with anti-IgM and anti-IgT Abs. Numbers in outlined boxes indicate the percentages of IgM + (top left) and IgT + (bottom right) B cells in the lymphocyte gate, respectively. ( E ) Frequency of IgM + and IgT + cells among total B cells present in trout pharynx and head kidney. ( F and G ) Immunoblot and densitometric analysis of the concentration of IgT, IgM, and IgD in pharyngeal mucus (F) and serum (G). ( H and I ) Ratio of IgT to IgM concentration (H) and IgD to IgM concentration (I) in pharyngeal mucus and serum, calculated from the values shown in (F) and (G), respectively. Results in (E)–(I) are expressed as mean ± SEM obtained from 12 individual fishes. MP, muscularis propria; PE, pharyngeal epithelium; SB, stratum basale; SM, submucosa.

    Journal: The Journal of Immunology Author Choice

    Article Title: Pharyngeal Immunity in Early Vertebrates Provides Functional and Evolutionary Insight into Mucosal Homeostasis

    doi: 10.4049/jimmunol.1900863

    Figure Lengend Snippet: General organization of teleost PM lymphoid tissue. ( A ) H E stain of the pharynx of a control adult rainbow trout ( O. mykiss ). Black arrowhead indicates taste bud. ( B ) H E (upper) and Alcian blue (lower) stain of the pharynx from five different families of control adult teleost fish. From left to right: grass carp ( C. idellus ), southern catfish ( S. meridionalis ), mandarin fish ( S. chuatsi ), snakehead ( O. argus Cantor), and rainbow trout ( O. mykiss ). Scale bar, 50 μm. Black and red arrowheads indicate lymphocytes and mucus cells, respectively. ( C ) Heat map illustrates results from qPCR of mRNAs for selected immune markers in trout skin, gut, gill, pharynx, spleen, and head kidney ( n = 6). Data are expressed as mean cycle threshold (Ct) values ± SEM. ( D ) Flow cytometry analysis of pharynx (left) and head kidney (right) leukocytes stained with anti-IgM and anti-IgT Abs. Numbers in outlined boxes indicate the percentages of IgM + (top left) and IgT + (bottom right) B cells in the lymphocyte gate, respectively. ( E ) Frequency of IgM + and IgT + cells among total B cells present in trout pharynx and head kidney. ( F and G ) Immunoblot and densitometric analysis of the concentration of IgT, IgM, and IgD in pharyngeal mucus (F) and serum (G). ( H and I ) Ratio of IgT to IgM concentration (H) and IgD to IgM concentration (I) in pharyngeal mucus and serum, calculated from the values shown in (F) and (G), respectively. Results in (E)–(I) are expressed as mean ± SEM obtained from 12 individual fishes. MP, muscularis propria; PE, pharyngeal epithelium; SB, stratum basale; SM, submucosa.

    Article Snippet: After 24 h, leukocytes from pharyngeal tissue or head kidney were obtained as described above, and cells were incubated with mAb mouse anti-trout IgM or anti-trout IgT (1 μg/ml each) on ice for 1 h. After washing three times, Alexa Fluor 488 goat anti-mouse IgG1 or Alexa Fluor 488 goat anti-mouse IgG2b (3 μg/ml each; Invitrogen) were used as secondary Ab to detect IgM+ or IgT+ B cells, respectively.

    Techniques: Staining, Fluorescence In Situ Hybridization, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry, Concentration Assay

    Local IgT-, IgM-, and IgD-specific responses to I. multifiliis parasite of survivor fish. Pharynx, head kidney, and spleen explants (∼30 mg each) from control and survivor fish were cultured in medium (400 μl) for 7 d. ( A – C ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in the culture medium of pharynx (A), head kidney (B), and spleen (C) (dilution 1:2) from control and survivor fish. ( D – F ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of pharynx (D), head kidney (E), and spleen (F) culture medium from survivor fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control fish ( n = 12). Data are representative of at least three independent experiments (mean ± SEM). * p

    Journal: The Journal of Immunology Author Choice

    Article Title: Pharyngeal Immunity in Early Vertebrates Provides Functional and Evolutionary Insight into Mucosal Homeostasis

    doi: 10.4049/jimmunol.1900863

    Figure Lengend Snippet: Local IgT-, IgM-, and IgD-specific responses to I. multifiliis parasite of survivor fish. Pharynx, head kidney, and spleen explants (∼30 mg each) from control and survivor fish were cultured in medium (400 μl) for 7 d. ( A – C ) Western blot analysis of IgT-, IgM-, and IgD-specific binding to I. multifiliis in the culture medium of pharynx (A), head kidney (B), and spleen (C) (dilution 1:2) from control and survivor fish. ( D – F ) IgT-, IgM-, and IgD-specific binding to I. multifiliis in dilutions of pharynx (D), head kidney (E), and spleen (F) culture medium from survivor fish, evaluated by densitometric analysis of immunoblots and presented as relative values to those of control fish ( n = 12). Data are representative of at least three independent experiments (mean ± SEM). * p

    Article Snippet: After 24 h, leukocytes from pharyngeal tissue or head kidney were obtained as described above, and cells were incubated with mAb mouse anti-trout IgM or anti-trout IgT (1 μg/ml each) on ice for 1 h. After washing three times, Alexa Fluor 488 goat anti-mouse IgG1 or Alexa Fluor 488 goat anti-mouse IgG2b (3 μg/ml each; Invitrogen) were used as secondary Ab to detect IgM+ or IgT+ B cells, respectively.

    Techniques: Fluorescence In Situ Hybridization, Cell Culture, Western Blot, Binding Assay

    OmpC and OmpF from S . Typhi promote antibody responses to ovalbumin (OVA) . Four BALB/c mice per group were immunized i.p. with 100 μg of OVA, OVA + 10 μg of porins, OVA + 10 μg of PorK, OVA + 5 μg of LPS, or OVA + 100 μg of alum. (A) Total IgG antibody responses were measured at the indicated time points by ELISA (B) high-avidity IgG and (C) IgG1, IgG2a, and IgG2b were measured at day 30 post-immunization. (D) BALB/c mice were injected i.p. with a depleting anti-CD4 mAb (GK1.5) 3 days prior to immunization with OVA or OVA + porins. GK1.5 mAb was given every 3 days thereafter until day 20, and the serum IgG and IgM responses were measured by ELISA. Mean + SEM are shown. These data are representative of three independent experiments. Statistical analysis was performed using one-way ANOVA with Bonferroni test post hoc . For panel (A) , Student’s t -test was performed comparing only OVA versus OVA + porins groups. Statistical differences are depicted as * P

    Journal: Frontiers in Immunology

    Article Title: Salmonella Typhi Porins OmpC and OmpF Are Potent Adjuvants for T-Dependent and T-Independent Antigens

    doi: 10.3389/fimmu.2017.00230

    Figure Lengend Snippet: OmpC and OmpF from S . Typhi promote antibody responses to ovalbumin (OVA) . Four BALB/c mice per group were immunized i.p. with 100 μg of OVA, OVA + 10 μg of porins, OVA + 10 μg of PorK, OVA + 5 μg of LPS, or OVA + 100 μg of alum. (A) Total IgG antibody responses were measured at the indicated time points by ELISA (B) high-avidity IgG and (C) IgG1, IgG2a, and IgG2b were measured at day 30 post-immunization. (D) BALB/c mice were injected i.p. with a depleting anti-CD4 mAb (GK1.5) 3 days prior to immunization with OVA or OVA + porins. GK1.5 mAb was given every 3 days thereafter until day 20, and the serum IgG and IgM responses were measured by ELISA. Mean + SEM are shown. These data are representative of three independent experiments. Statistical analysis was performed using one-way ANOVA with Bonferroni test post hoc . For panel (A) , Student’s t -test was performed comparing only OVA versus OVA + porins groups. Statistical differences are depicted as * P

    Article Snippet: Peroxidase-conjugated anti-mouse IgM, IgG H + L, IgG2a, IgG2b, or IgG3 (Invitrogen, CA, USA) were diluted at a ratio of 1:1,000.

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

    S . Typhi porins promote antibody responses to the T-independent anti-typhoid Vi antigen . Four BALB/c mice per group were immunized i.p. with 10 μg of Vi CPS vaccine (Typhim), Vi + 10 μg porins or Vi + 10 μg of PorK on days 0 and 15. (A) IgM and total IgG responses and (B) IgG1 and IgG3 responses against Vi CPS were measured by ELISA on day 20 post-immunization. Mean + SEM are plotted. These data are representative of two independent experiments. Statistical analysis was performed using one-way ANOVA with Bonferroni test post hoc . Statistical differences are depicted as * P

    Journal: Frontiers in Immunology

    Article Title: Salmonella Typhi Porins OmpC and OmpF Are Potent Adjuvants for T-Dependent and T-Independent Antigens

    doi: 10.3389/fimmu.2017.00230

    Figure Lengend Snippet: S . Typhi porins promote antibody responses to the T-independent anti-typhoid Vi antigen . Four BALB/c mice per group were immunized i.p. with 10 μg of Vi CPS vaccine (Typhim), Vi + 10 μg porins or Vi + 10 μg of PorK on days 0 and 15. (A) IgM and total IgG responses and (B) IgG1 and IgG3 responses against Vi CPS were measured by ELISA on day 20 post-immunization. Mean + SEM are plotted. These data are representative of two independent experiments. Statistical analysis was performed using one-way ANOVA with Bonferroni test post hoc . Statistical differences are depicted as * P

    Article Snippet: Peroxidase-conjugated anti-mouse IgM, IgG H + L, IgG2a, IgG2b, or IgG3 (Invitrogen, CA, USA) were diluted at a ratio of 1:1,000.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    The production of IL-10 by PBMC or CD8-depleted cells (– CD8) from individuals with Graves’ disease or Hashimoto’s thyroiditis, and normal controls. The PBMC or CD8-depleted cells were incubated in the presence or absence of anti-CD40 MoAb and IL-4. * P

    Journal: Clinical and Experimental Immunology

    Article Title: The effects of CD40- and interleukin (IL-4)-activated CD23+ cells on the production of IL-10 by mononuclear cells in Graves' disease: the role of CD8+ cells

    doi: 10.1046/j.1365-2249.2002.01818.x

    Figure Lengend Snippet: The production of IL-10 by PBMC or CD8-depleted cells (– CD8) from individuals with Graves’ disease or Hashimoto’s thyroiditis, and normal controls. The PBMC or CD8-depleted cells were incubated in the presence or absence of anti-CD40 MoAb and IL-4. * P

    Article Snippet: Peripheral blood mononuclear cells (PBMC) were separated through a Ficoll-Paque (Pharmacia, Uppsala, Sweden) density gradient and cultured for 7 days at 37°C at 1 × 106 cells/ml in RPMI-1640 medium containing 10% fetal bovine serum (FBS) with 2 μg/ml anti-CD40 monoclonal antibody (mouse monoclonal IgM antibody to human CD40, clone 14G7, Caltag Laboratory, San Francisco, CA, USA) and human recombinant interleukin-4 (Final conc.

    Techniques: Incubation

    The production of soluble CD23 by PBMC or CD8-depleted cells (– CD8) from individuals with Graves’ disease or Hashimoto’s thyroiditis, and normal controls. The PBMC or CD8-depleted cells were incubated in the presence or absence of anti-CD40 MoAb and IL-4. * P

    Journal: Clinical and Experimental Immunology

    Article Title: The effects of CD40- and interleukin (IL-4)-activated CD23+ cells on the production of IL-10 by mononuclear cells in Graves' disease: the role of CD8+ cells

    doi: 10.1046/j.1365-2249.2002.01818.x

    Figure Lengend Snippet: The production of soluble CD23 by PBMC or CD8-depleted cells (– CD8) from individuals with Graves’ disease or Hashimoto’s thyroiditis, and normal controls. The PBMC or CD8-depleted cells were incubated in the presence or absence of anti-CD40 MoAb and IL-4. * P

    Article Snippet: Peripheral blood mononuclear cells (PBMC) were separated through a Ficoll-Paque (Pharmacia, Uppsala, Sweden) density gradient and cultured for 7 days at 37°C at 1 × 106 cells/ml in RPMI-1640 medium containing 10% fetal bovine serum (FBS) with 2 μg/ml anti-CD40 monoclonal antibody (mouse monoclonal IgM antibody to human CD40, clone 14G7, Caltag Laboratory, San Francisco, CA, USA) and human recombinant interleukin-4 (Final conc.

    Techniques: Incubation

    The percentages of CD23 + or CD20 + cells in PBMC or CD8-depleted cells (– CD8) from individuals with Graves’ disease or Hashimoto’s thyroiditis, and normal controls. The PBMC or CD8-depleted cells were incubated in the presence or absence of anti-CD40 mAb and IL-4. * P

    Journal: Clinical and Experimental Immunology

    Article Title: The effects of CD40- and interleukin (IL-4)-activated CD23+ cells on the production of IL-10 by mononuclear cells in Graves' disease: the role of CD8+ cells

    doi: 10.1046/j.1365-2249.2002.01818.x

    Figure Lengend Snippet: The percentages of CD23 + or CD20 + cells in PBMC or CD8-depleted cells (– CD8) from individuals with Graves’ disease or Hashimoto’s thyroiditis, and normal controls. The PBMC or CD8-depleted cells were incubated in the presence or absence of anti-CD40 mAb and IL-4. * P

    Article Snippet: Peripheral blood mononuclear cells (PBMC) were separated through a Ficoll-Paque (Pharmacia, Uppsala, Sweden) density gradient and cultured for 7 days at 37°C at 1 × 106 cells/ml in RPMI-1640 medium containing 10% fetal bovine serum (FBS) with 2 μg/ml anti-CD40 monoclonal antibody (mouse monoclonal IgM antibody to human CD40, clone 14G7, Caltag Laboratory, San Francisco, CA, USA) and human recombinant interleukin-4 (Final conc.

    Techniques: Incubation