goat anti mouse secondary antibody  (Thermo Fisher)


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    Name:
    Goat anti Mouse IgA Secondary Antibody
    Description:
    Goat anti Mouse IgA Secondary Antibody for Western Blot IHC ELISA
    Catalog Number:
    626722
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Antibodies and Secondary Detection|Cell Analysis|Secondary Detection
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    Structured Review

    Thermo Fisher goat anti mouse secondary antibody
    Endophilin A2 knockout <t>mouse</t> mimics the phenotype of CRISPR-targeted chimeras and exhibits reduced levels of affinity matured serum <t>antibodies</t> upon immunization. (A) Characterization of B cell populations in spleen, bone marrow and lymph node at steady state. Data show mean and SEM from N = 5 mice. P, significance in t tests performed for follicular (FO) and MZ cell counts. (B) Spleen size in Sh3gl1 -/- and WT littermates at 12 weeks of age. (C) Surface levels of MHCII, BAFFR, CD40 and LPAM1 in Sh3gl1 -/- and WT littermates. P, statistical significance from unpaired t tests. (D) Quantification of Fas + CD38 - NP-specific GC population as percentage of total splenic B cells 14 days after NP-CGG immunization in alum. N = 6 mice. (E-F) Serum antibodies collected pre-immunization and 7- or 14-days post NP-CGG immunization, detected by NP7-BSA ELISA using isotypespecific <t>secondary</t> antibodies. Data show mean ± SEM from N = 3 experiments with statistical significance from 2-way ANOVA with multiple comparisons. (G) Ratio of binding to NP7 and NP25, as measured by ELISA. (H) Numbers of Tfh cells 14 days post-immunization, identified as a CD4 + CD44 + PD-1 + CXCR5 + population. Mean and SEM of N = 4 mice.
    Goat anti Mouse IgA Secondary Antibody for Western Blot IHC ELISA
    https://www.bioz.com/result/goat anti mouse secondary antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse secondary antibody - by Bioz Stars, 2021-04
    97/100 stars

    Images

    1) 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 knockout mouse mimics the phenotype of CRISPR-targeted chimeras and exhibits reduced levels of affinity matured serum antibodies upon immunization. (A) Characterization of B cell populations in spleen, bone marrow and lymph node at steady state. Data show mean and SEM from N = 5 mice. P, significance in t tests performed for follicular (FO) and MZ cell counts. (B) Spleen size in Sh3gl1 -/- and WT littermates at 12 weeks of age. (C) Surface levels of MHCII, BAFFR, CD40 and LPAM1 in Sh3gl1 -/- and WT littermates. P, statistical significance from unpaired t tests. (D) Quantification of Fas + CD38 - NP-specific GC population as percentage of total splenic B cells 14 days after NP-CGG immunization in alum. N = 6 mice. (E-F) Serum antibodies collected pre-immunization and 7- or 14-days post NP-CGG immunization, detected by NP7-BSA ELISA using isotypespecific secondary antibodies. Data show mean ± SEM from N = 3 experiments with statistical significance from 2-way ANOVA with multiple comparisons. (G) Ratio of binding to NP7 and NP25, as measured by ELISA. (H) Numbers of Tfh cells 14 days post-immunization, identified as a CD4 + CD44 + PD-1 + CXCR5 + population. Mean and SEM of N = 4 mice.
    Figure Legend Snippet: Endophilin A2 knockout mouse mimics the phenotype of CRISPR-targeted chimeras and exhibits reduced levels of affinity matured serum antibodies upon immunization. (A) Characterization of B cell populations in spleen, bone marrow and lymph node at steady state. Data show mean and SEM from N = 5 mice. P, significance in t tests performed for follicular (FO) and MZ cell counts. (B) Spleen size in Sh3gl1 -/- and WT littermates at 12 weeks of age. (C) Surface levels of MHCII, BAFFR, CD40 and LPAM1 in Sh3gl1 -/- and WT littermates. P, statistical significance from unpaired t tests. (D) Quantification of Fas + CD38 - NP-specific GC population as percentage of total splenic B cells 14 days after NP-CGG immunization in alum. N = 6 mice. (E-F) Serum antibodies collected pre-immunization and 7- or 14-days post NP-CGG immunization, detected by NP7-BSA ELISA using isotypespecific secondary antibodies. Data show mean ± SEM from N = 3 experiments with statistical significance from 2-way ANOVA with multiple comparisons. (G) Ratio of binding to NP7 and NP25, as measured by ELISA. (H) Numbers of Tfh cells 14 days post-immunization, identified as a CD4 + CD44 + PD-1 + CXCR5 + population. Mean and SEM of N = 4 mice.

    Techniques Used: Knock-Out, CRISPR, Mouse Assay, Enzyme-linked Immunosorbent Assay, Binding Assay

    2) Product Images from "Functional insights of two MATE transporters from Vibrio fluvialis"

    Article Title: Functional insights of two MATE transporters from Vibrio fluvialis

    Journal: bioRxiv

    doi: 10.1101/2021.01.02.425065

    Membrane localisation of recombinant efflux pumps in E. coli. Panel A. Confocal imaging of pBAD-recombinants in heterologous E. coli host. A/D :Visualisation of E. coli cells harbouring pBAD- vfh/vfd stained with membrane marker Film-tracer 1-43; B/E :Detection of recombinant pBAD- vfh/vfd in E. coli labelled with Goat anti-Mouse Cy3 secondary antibody; C/F (63X ZOOM 4): Superimposed/merged images from A/B or D/E of recombinant pBAD- vfh/vfd in E. coli LMG194; G/J : Visualisation of E. coli cells harbouring pBAD- vfh/vfd using membrane marker Film-tracer 1-43; H/K : Detection of recombinant pBAD- vfh/vfd in E. coli labelled with Goat anti-Mouse Cy3 secondary antibody; I/L : Superimposed/merged images from G/H or J/K of recombinant pBAD- vfh/vfd in E. coli K-12 Δ tolC 2 Δ tolC . Panel B. Membrane fractionation of E. coli harbouring pBAD- vfd. 63X Zoom 4 settings were used for obtaining these images. M- Spectramulticolour protein marker (Fermentas). Lane 1- VFD −ve- Total cell lysates for uninduced samples; Lane 2-VFD +ve- Total cell lysates for induced samples; Lane 3- VFD Son (Sup) - total soluble fraction after sonication.; Lane 4- VFD SAR (Sup)- Total soluble proteins after sarkosyl treatment; and Lane 5- VFD SAR Pellet- debris and other subcellular organelles. Lane 6- Lac-HisPositive control. Goat anti-V5 antibody was used as primary antibody. HRP conjugated Mouse Anti-Goat IgG was used as secondary antibody (Jackson Immunoresearch). Prestained Spectramulticolour (Fermentas) molecular weight marker in kDa has been indicated of the left of panel. The blots were developed with Novex ECL substrates (Invitrogen).
    Figure Legend Snippet: Membrane localisation of recombinant efflux pumps in E. coli. Panel A. Confocal imaging of pBAD-recombinants in heterologous E. coli host. A/D :Visualisation of E. coli cells harbouring pBAD- vfh/vfd stained with membrane marker Film-tracer 1-43; B/E :Detection of recombinant pBAD- vfh/vfd in E. coli labelled with Goat anti-Mouse Cy3 secondary antibody; C/F (63X ZOOM 4): Superimposed/merged images from A/B or D/E of recombinant pBAD- vfh/vfd in E. coli LMG194; G/J : Visualisation of E. coli cells harbouring pBAD- vfh/vfd using membrane marker Film-tracer 1-43; H/K : Detection of recombinant pBAD- vfh/vfd in E. coli labelled with Goat anti-Mouse Cy3 secondary antibody; I/L : Superimposed/merged images from G/H or J/K of recombinant pBAD- vfh/vfd in E. coli K-12 Δ tolC 2 Δ tolC . Panel B. Membrane fractionation of E. coli harbouring pBAD- vfd. 63X Zoom 4 settings were used for obtaining these images. M- Spectramulticolour protein marker (Fermentas). Lane 1- VFD −ve- Total cell lysates for uninduced samples; Lane 2-VFD +ve- Total cell lysates for induced samples; Lane 3- VFD Son (Sup) - total soluble fraction after sonication.; Lane 4- VFD SAR (Sup)- Total soluble proteins after sarkosyl treatment; and Lane 5- VFD SAR Pellet- debris and other subcellular organelles. Lane 6- Lac-HisPositive control. Goat anti-V5 antibody was used as primary antibody. HRP conjugated Mouse Anti-Goat IgG was used as secondary antibody (Jackson Immunoresearch). Prestained Spectramulticolour (Fermentas) molecular weight marker in kDa has been indicated of the left of panel. The blots were developed with Novex ECL substrates (Invitrogen).

    Techniques Used: Recombinant, Imaging, Staining, Marker, Fractionation, Sonication, Molecular Weight

    Related Articles

    Expressing:

    Article Title: Structural basis of prostate-specific membrane antigen recognition by the A9g RNA aptamer
    Article Snippet: Data were processed in FlowJo software (FlowJo, LLC, Ashland, OR, USA). .. To analyze surface expression levels of PSMA mutants, the 5D3 monoclonal antibody ( ) was used at 40 nM concentration and incubated with cells on ice for 20 min. After washing with PBS supplemented with 0.5% (w/v) gelatin, cells were labeled with goat anti-mouse secondary antibody conjugated with Alexa Fluor647 (Invitrogen, Carlsbad, CA; 4 μg/ml) on ice for 20 min. Further washing, labeling, flow cytometry and analysis steps were performed as described above. ..

    Concentration Assay:

    Article Title: Structural basis of prostate-specific membrane antigen recognition by the A9g RNA aptamer
    Article Snippet: Data were processed in FlowJo software (FlowJo, LLC, Ashland, OR, USA). .. To analyze surface expression levels of PSMA mutants, the 5D3 monoclonal antibody ( ) was used at 40 nM concentration and incubated with cells on ice for 20 min. After washing with PBS supplemented with 0.5% (w/v) gelatin, cells were labeled with goat anti-mouse secondary antibody conjugated with Alexa Fluor647 (Invitrogen, Carlsbad, CA; 4 μg/ml) on ice for 20 min. Further washing, labeling, flow cytometry and analysis steps were performed as described above. ..

    Incubation:

    Article Title: Structural basis of prostate-specific membrane antigen recognition by the A9g RNA aptamer
    Article Snippet: Data were processed in FlowJo software (FlowJo, LLC, Ashland, OR, USA). .. To analyze surface expression levels of PSMA mutants, the 5D3 monoclonal antibody ( ) was used at 40 nM concentration and incubated with cells on ice for 20 min. After washing with PBS supplemented with 0.5% (w/v) gelatin, cells were labeled with goat anti-mouse secondary antibody conjugated with Alexa Fluor647 (Invitrogen, Carlsbad, CA; 4 μg/ml) on ice for 20 min. Further washing, labeling, flow cytometry and analysis steps were performed as described above. ..

    Article Title: Mouse glutamate carboxypeptidase  II ( GCPII) has a similar enzyme activity and inhibition profile but a different tissue distribution to human GCPII
    Article Snippet: .. After blotting, the membrane was blocked with 0.55% (w/v) casein solution in PBS (Casein Buffer 20X‐4X Concentrate, SDT, Baesweiler, Germany) at room temperature for 1 h. To visualize GCPII, the blots were probed with the antibody GCP‐04 (described in ) for 12 h at 4 °C (200 ng·mL−1 ; diluted in 0.55% casein solution), washed three times with PBS containing 0.05% Tween 20 (PBST buffer), and incubated with goat anti‐mouse antibody conjugated with horseradish peroxidase (Thermo Scientific, Waltham, MA, USA; diluted in 0.55% casein solution, 1 : 25 000). .. The blots were then washed three times with PBST to remove unbound antibodies and developed with SuperSignal West Femto Chemiluminescent Substrate (Thermo Scientific).

    Article Title: A Primate APOL1 Variant That Kills Trypanosoma brucei gambiense
    Article Snippet: After washing three times in PBS, slides were incubated for 40 minutes with 1:500 mouse anti-p67 antibody (gift from Jay Bangs, Department of Microbiology and Immunology, University at Buffalo, NY, USA) in blocking solution. .. Washes were repeated and then primary antibody was detected using 1:1000 goat anti-mouse AlexaFluor594 secondary antibody (Life technologies) incubated for 40 minutes in blocking solution. .. To detect His-tagged APOL1 slides were washed three times in PBS and then incubated for 40 minutes with 1:500 AlexaFluor488 mouse anti-penta-His antibody (Molecular Probes, Invitrogen) in blocking solution.

    Article Title: The S1/S2 boundary of SARS-CoV-2 spike protein modulates cell entry pathways and transmission
    Article Snippet: Immunofluorescence assay Virus-infected cells were washed twice with PBS, fixed with 4% paraformaldehyde in PBS for 30 min, permeablized with 0.2% Triton X-100 for 1 h. Cells were then incubated with house-made mouse anti-SARS-CoV-2 nucleocapsid protein serum (1:1000) at 4 °C overnight. .. After three washes, cells were incubated with the secondary goat anti-mouse antibody conjugated with Alexa Fluor 555 (Thermo #A-21424, 2 μg/ml) for 2 h at room temperature, followed by staining with 4’,6-diamidino-2-phenylindole (DAPI). .. Images were collected using an Operetta High Content Imaging System (PerkinElmer), and processed using the ImageJ program ( http://rsb.info.nih.gov/ij/).

    Article Title: Ribavirin efficiently suppresses porcine nidovirus replication
    Article Snippet: .. The cells were blocked with 1% bovine serum albumin (BSA) in PBS for 30 min at RT and then incubated with N-specific MAb 7 for 2 h. After being washed five times in PBS, the cells were incubated for 1 h at RT with a goat anti-mouse secondary antibody conjugated with Alexa Fluor 488 (Molecular Probes, Carlsbad, CA), followed by counterstaining with 4′,6-diamidino-2-phenylindole (DAPI; Sigma). .. The coverslips were mounted on microscope glass slides in mounting buffer (60% glycerol and 0.1% sodium azide in PBS) and cell staining was visualized using a fluorescent Leica DM IL LED microscope (Leica, Wetzlar, Germany).

    Article Title: Comparative analysis reveals the species-specific genetic determinants of ACE2 required for SARS-CoV-2 entry
    Article Snippet: Analysis of SARS-CoV-2 infection by high-content imaging systemA549 cells were transduced with lentiviruses expressing the ACE2 of different species for 48 h. Cells were then infected with nCoV-SH01 (SARS-CoV-2) at an MOI of 1 for 1 h, washed three times with PBS, and incubated in 2% FBS culture medium for 48 h. Cell were then fixed for viral antigen staining with 4% paraformaldehyde in PBS, permeablized with 0.2% Triton X-100, and incubated with a rabbit polyclonal antibody against the SARS-CoV nucleocapsid protein (Rockland, 200-401-A50, 1μg/ml) and a mouse anti-FLAG M2 antibody (Sigma-Aldrich #1804, 1μg/ml) at 4 °C overnight. .. After three washes, cells were incubated with a secondary goat anti-rabbit antibody conjugated with Alexa Fluor 555 (Thermo #A32732, 2 μg/ml) and goat anti-mouse antibody conjugated with Alexa Fluor 647 (Thermo #A21235, 2 μg/ml) for 2 h at room temperature, followed by staining with 4’,6-diamidino-2-phenylindole (DAPI). .. Images were collected using an Operetta High-Content Imaging System (PerkinElmer).

    Labeling:

    Article Title: Structural basis of prostate-specific membrane antigen recognition by the A9g RNA aptamer
    Article Snippet: Data were processed in FlowJo software (FlowJo, LLC, Ashland, OR, USA). .. To analyze surface expression levels of PSMA mutants, the 5D3 monoclonal antibody ( ) was used at 40 nM concentration and incubated with cells on ice for 20 min. After washing with PBS supplemented with 0.5% (w/v) gelatin, cells were labeled with goat anti-mouse secondary antibody conjugated with Alexa Fluor647 (Invitrogen, Carlsbad, CA; 4 μg/ml) on ice for 20 min. Further washing, labeling, flow cytometry and analysis steps were performed as described above. ..

    Flow Cytometry:

    Article Title: Structural basis of prostate-specific membrane antigen recognition by the A9g RNA aptamer
    Article Snippet: Data were processed in FlowJo software (FlowJo, LLC, Ashland, OR, USA). .. To analyze surface expression levels of PSMA mutants, the 5D3 monoclonal antibody ( ) was used at 40 nM concentration and incubated with cells on ice for 20 min. After washing with PBS supplemented with 0.5% (w/v) gelatin, cells were labeled with goat anti-mouse secondary antibody conjugated with Alexa Fluor647 (Invitrogen, Carlsbad, CA; 4 μg/ml) on ice for 20 min. Further washing, labeling, flow cytometry and analysis steps were performed as described above. ..

    Blocking Assay:

    Article Title: A Primate APOL1 Variant That Kills Trypanosoma brucei gambiense
    Article Snippet: After washing three times in PBS, slides were incubated for 40 minutes with 1:500 mouse anti-p67 antibody (gift from Jay Bangs, Department of Microbiology and Immunology, University at Buffalo, NY, USA) in blocking solution. .. Washes were repeated and then primary antibody was detected using 1:1000 goat anti-mouse AlexaFluor594 secondary antibody (Life technologies) incubated for 40 minutes in blocking solution. .. To detect His-tagged APOL1 slides were washed three times in PBS and then incubated for 40 minutes with 1:500 AlexaFluor488 mouse anti-penta-His antibody (Molecular Probes, Invitrogen) in blocking solution.

    Staining:

    Article Title: The S1/S2 boundary of SARS-CoV-2 spike protein modulates cell entry pathways and transmission
    Article Snippet: Immunofluorescence assay Virus-infected cells were washed twice with PBS, fixed with 4% paraformaldehyde in PBS for 30 min, permeablized with 0.2% Triton X-100 for 1 h. Cells were then incubated with house-made mouse anti-SARS-CoV-2 nucleocapsid protein serum (1:1000) at 4 °C overnight. .. After three washes, cells were incubated with the secondary goat anti-mouse antibody conjugated with Alexa Fluor 555 (Thermo #A-21424, 2 μg/ml) for 2 h at room temperature, followed by staining with 4’,6-diamidino-2-phenylindole (DAPI). .. Images were collected using an Operetta High Content Imaging System (PerkinElmer), and processed using the ImageJ program ( http://rsb.info.nih.gov/ij/).

    Article Title: Comparative analysis reveals the species-specific genetic determinants of ACE2 required for SARS-CoV-2 entry
    Article Snippet: Analysis of SARS-CoV-2 infection by high-content imaging systemA549 cells were transduced with lentiviruses expressing the ACE2 of different species for 48 h. Cells were then infected with nCoV-SH01 (SARS-CoV-2) at an MOI of 1 for 1 h, washed three times with PBS, and incubated in 2% FBS culture medium for 48 h. Cell were then fixed for viral antigen staining with 4% paraformaldehyde in PBS, permeablized with 0.2% Triton X-100, and incubated with a rabbit polyclonal antibody against the SARS-CoV nucleocapsid protein (Rockland, 200-401-A50, 1μg/ml) and a mouse anti-FLAG M2 antibody (Sigma-Aldrich #1804, 1μg/ml) at 4 °C overnight. .. After three washes, cells were incubated with a secondary goat anti-rabbit antibody conjugated with Alexa Fluor 555 (Thermo #A32732, 2 μg/ml) and goat anti-mouse antibody conjugated with Alexa Fluor 647 (Thermo #A21235, 2 μg/ml) for 2 h at room temperature, followed by staining with 4’,6-diamidino-2-phenylindole (DAPI). .. Images were collected using an Operetta High-Content Imaging System (PerkinElmer).

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  • 99
    Thermo Fisher anti igm
    Phenotypic and functional characterization of the IL-33-induced Breg-like cells (Breg IL-33 ) in the blood. A–C. Surface phenotype of Breg IL-33 : The IL-33-induced IL-10 producing B cells expressed high surface <t>IgM</t> (A), CD1d and CD25 (B), but down-regulated CD23 (FceRII) (B, C). Data shown in the table under (B) were results showing the Mean MFI (±SD) values calculated from results of 5 repeated experiments, as ratio of the IL-10 − B, and IL-10 + B, cell subset over the total B cells gated. Statistical analysis: p values (Student t test). D–F. In vitro Breg suppression assays: Immunosuppressive effects of the IL-33-induced Breg-like cells on B effector (Beff) cell proliferation (D), division (E), and its IL-10 dependency (F). D. CD23 + B cells (Beff, 10 5 ) were cultured in the presence of <t>anti-CD40</t> (2.5 μg/ml), with titrating (as indicated) doses of CD23 − B cells (Breg IL-33 ) purified from the IL-33-injected mice. Cell proliferation was determined by thymidine incorporation at 48 h s. E. Breg IL-33 (CD23 - ) purified from hIL-33-injected WT mice (Wk-2) were primed with anti-CD40 (2.5 μg/ml) for 5 h before being added to CFSE-labelled CD23 + Beff cells. Cell division (CFSE dilution) was determined by flow cytometry at day 5 of culture in the presence or absence of LPS (0.5 μg/ml) or anti-IgM (5 μg/ml). F. IL-10 levels in the culture supernatants were quantified by ELISA (BD Biosciences).
    Anti Igm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti igm/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti igm - by Bioz Stars, 2021-04
    99/100 stars
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    95
    Thermo Fisher igg2a
    Anti-M2e antibody response in BAL. BALB/c mice (n = 5/group) were immunised i.n. with 50 μg of Flg-2M2eh2M2ek recombinant protein on days 0, 14, 28. Mice of control group were administered with PBS. Two weeks post-second boost M2e-specific <t>IgG</t> (A) and sIgA (B) responses were evaluated by ELISA to M2eh and M2ek synthetic peptides. Horizontal bars indicate mean titres among 5 mice per group. Statistical significance was determined using the Mann-Whitney U-test. The P values between immunised and control groups are indicated.
    Igg2a, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg2a/product/Thermo Fisher
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    86
    Thermo Fisher alkaline phosphatase conjugated goat anti mouse igg
    Plasmodium falciparum EBA175RIII–V construct and expressed antigen. A schematic representation of the 1620 bp region of EBA175RIII–V cloned into the pLEA2 expression vector, which contains the nucleotide sequence of a hexahistidine tag inframe of the multiple cloning site ( a ). The culture supernatant (1) containing the secreted protein as well as the purified protein (2) was analysed by SDS-PAGE followed by coomassie staining ( b ) and a western blot probed with penta-His mouse monoclonal antibody <t>(IgG1)</t> ( c )
    Alkaline Phosphatase Conjugated Goat Anti Mouse Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alkaline phosphatase conjugated goat anti mouse igg/product/Thermo Fisher
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    Image Search Results


    Phenotypic and functional characterization of the IL-33-induced Breg-like cells (Breg IL-33 ) in the blood. A–C. Surface phenotype of Breg IL-33 : The IL-33-induced IL-10 producing B cells expressed high surface IgM (A), CD1d and CD25 (B), but down-regulated CD23 (FceRII) (B, C). Data shown in the table under (B) were results showing the Mean MFI (±SD) values calculated from results of 5 repeated experiments, as ratio of the IL-10 − B, and IL-10 + B, cell subset over the total B cells gated. Statistical analysis: p values (Student t test). D–F. In vitro Breg suppression assays: Immunosuppressive effects of the IL-33-induced Breg-like cells on B effector (Beff) cell proliferation (D), division (E), and its IL-10 dependency (F). D. CD23 + B cells (Beff, 10 5 ) were cultured in the presence of anti-CD40 (2.5 μg/ml), with titrating (as indicated) doses of CD23 − B cells (Breg IL-33 ) purified from the IL-33-injected mice. Cell proliferation was determined by thymidine incorporation at 48 h s. E. Breg IL-33 (CD23 - ) purified from hIL-33-injected WT mice (Wk-2) were primed with anti-CD40 (2.5 μg/ml) for 5 h before being added to CFSE-labelled CD23 + Beff cells. Cell division (CFSE dilution) was determined by flow cytometry at day 5 of culture in the presence or absence of LPS (0.5 μg/ml) or anti-IgM (5 μg/ml). F. IL-10 levels in the culture supernatants were quantified by ELISA (BD Biosciences).

    Journal: Journal of Autoimmunity

    Article Title: IL-10-producing regulatory B cells induced by IL-33 (BregIL-33) effectively attenuate mucosal inflammatory responses in the gut

    doi: 10.1016/j.jaut.2014.01.032

    Figure Lengend Snippet: Phenotypic and functional characterization of the IL-33-induced Breg-like cells (Breg IL-33 ) in the blood. A–C. Surface phenotype of Breg IL-33 : The IL-33-induced IL-10 producing B cells expressed high surface IgM (A), CD1d and CD25 (B), but down-regulated CD23 (FceRII) (B, C). Data shown in the table under (B) were results showing the Mean MFI (±SD) values calculated from results of 5 repeated experiments, as ratio of the IL-10 − B, and IL-10 + B, cell subset over the total B cells gated. Statistical analysis: p values (Student t test). D–F. In vitro Breg suppression assays: Immunosuppressive effects of the IL-33-induced Breg-like cells on B effector (Beff) cell proliferation (D), division (E), and its IL-10 dependency (F). D. CD23 + B cells (Beff, 10 5 ) were cultured in the presence of anti-CD40 (2.5 μg/ml), with titrating (as indicated) doses of CD23 − B cells (Breg IL-33 ) purified from the IL-33-injected mice. Cell proliferation was determined by thymidine incorporation at 48 h s. E. Breg IL-33 (CD23 - ) purified from hIL-33-injected WT mice (Wk-2) were primed with anti-CD40 (2.5 μg/ml) for 5 h before being added to CFSE-labelled CD23 + Beff cells. Cell division (CFSE dilution) was determined by flow cytometry at day 5 of culture in the presence or absence of LPS (0.5 μg/ml) or anti-IgM (5 μg/ml). F. IL-10 levels in the culture supernatants were quantified by ELISA (BD Biosciences).

    Article Snippet: Respectively, fixed numbers (105 ) of B responder cells (CD19+ CD23+ ) were cultured, in the presence or absence of anti-CD40 (2.5 μg/ml, Enzo Life Sciences, UK), anti-IgM (5 μg/ml, Thermo Scientific, UK) or LPS (0.5 μg/ml, Sigma–Aldrich, UK), with titrated doses of BregIL-33 (CD19+ CD23− ) isolated from IL-33-treated WT or IL-10−/− mice as described above.

    Techniques: Functional Assay, In Vitro, Cell Culture, Purification, Injection, Mouse Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    Anti-M2e antibody response in BAL. BALB/c mice (n = 5/group) were immunised i.n. with 50 μg of Flg-2M2eh2M2ek recombinant protein on days 0, 14, 28. Mice of control group were administered with PBS. Two weeks post-second boost M2e-specific IgG (A) and sIgA (B) responses were evaluated by ELISA to M2eh and M2ek synthetic peptides. Horizontal bars indicate mean titres among 5 mice per group. Statistical significance was determined using the Mann-Whitney U-test. The P values between immunised and control groups are indicated.

    Journal: PLoS ONE

    Article Title: Protection against Multiple Influenza A Virus Strains Induced by Candidate Recombinant Vaccine Based on Heterologous M2e Peptides Linked to Flagellin

    doi: 10.1371/journal.pone.0119520

    Figure Lengend Snippet: Anti-M2e antibody response in BAL. BALB/c mice (n = 5/group) were immunised i.n. with 50 μg of Flg-2M2eh2M2ek recombinant protein on days 0, 14, 28. Mice of control group were administered with PBS. Two weeks post-second boost M2e-specific IgG (A) and sIgA (B) responses were evaluated by ELISA to M2eh and M2ek synthetic peptides. Horizontal bars indicate mean titres among 5 mice per group. Statistical significance was determined using the Mann-Whitney U-test. The P values between immunised and control groups are indicated.

    Article Snippet: HRP-labelled goat anti-mouse mAb IgG (Invitrogen, USA), goat anti-mouse mAb IgG1 (Invitrogen, USA), IgG2a (Invitrogen, USA), and IgA (Invitrogen, USA) were diluted in PBS with 5% FCS and added (100 μl/well) into the plates.

    Techniques: Mouse Assay, Recombinant, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Anti-M2e antibody response in serum. BALB/c mice (n = 5/group) were immunised i.n. with 50 μg of Flg-2M2eh2M2ek recombinant protein on days 0, 14, 28. Mice of control group were administered with PBS. Two weeks post-first (A) and second boosts (B) M2e-specific IgG responses were evaluated by ELISA to M2eh and M2ek synthetic peptides. Horizontal bars indicate mean titres among 5 mice per group. (C) Anti-M2e IgG subclasses tested against M2ek and M2eh in serum 2 weeks post-second boost were determined by ELISA. Statistically significant differences between IgG1 and IgG2a levels: p

    Journal: PLoS ONE

    Article Title: Protection against Multiple Influenza A Virus Strains Induced by Candidate Recombinant Vaccine Based on Heterologous M2e Peptides Linked to Flagellin

    doi: 10.1371/journal.pone.0119520

    Figure Lengend Snippet: Anti-M2e antibody response in serum. BALB/c mice (n = 5/group) were immunised i.n. with 50 μg of Flg-2M2eh2M2ek recombinant protein on days 0, 14, 28. Mice of control group were administered with PBS. Two weeks post-first (A) and second boosts (B) M2e-specific IgG responses were evaluated by ELISA to M2eh and M2ek synthetic peptides. Horizontal bars indicate mean titres among 5 mice per group. (C) Anti-M2e IgG subclasses tested against M2ek and M2eh in serum 2 weeks post-second boost were determined by ELISA. Statistically significant differences between IgG1 and IgG2a levels: p

    Article Snippet: HRP-labelled goat anti-mouse mAb IgG (Invitrogen, USA), goat anti-mouse mAb IgG1 (Invitrogen, USA), IgG2a (Invitrogen, USA), and IgA (Invitrogen, USA) were diluted in PBS with 5% FCS and added (100 μl/well) into the plates.

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

    Plasmodium falciparum EBA175RIII–V construct and expressed antigen. A schematic representation of the 1620 bp region of EBA175RIII–V cloned into the pLEA2 expression vector, which contains the nucleotide sequence of a hexahistidine tag inframe of the multiple cloning site ( a ). The culture supernatant (1) containing the secreted protein as well as the purified protein (2) was analysed by SDS-PAGE followed by coomassie staining ( b ) and a western blot probed with penta-His mouse monoclonal antibody (IgG1) ( c )

    Journal: Malaria Journal

    Article Title: Assessment of the quality and quantity of naturally induced antibody responses to EBA175RIII–V in Ghanaian children living in two communities with varying malaria transmission patterns

    doi: 10.1186/s12936-017-2167-3

    Figure Lengend Snippet: Plasmodium falciparum EBA175RIII–V construct and expressed antigen. A schematic representation of the 1620 bp region of EBA175RIII–V cloned into the pLEA2 expression vector, which contains the nucleotide sequence of a hexahistidine tag inframe of the multiple cloning site ( a ). The culture supernatant (1) containing the secreted protein as well as the purified protein (2) was analysed by SDS-PAGE followed by coomassie staining ( b ) and a western blot probed with penta-His mouse monoclonal antibody (IgG1) ( c )

    Article Snippet: The western blot was probed with penta-His mouse IgG1 monoclonal antibodies (Thermo Scientific, USA) followed by alkaline phosphatase conjugated goat anti mouse IgG (H + L) secondary antibodies (Thermo Scientific, USA).

    Techniques: Construct, Clone Assay, Expressing, Plasmid Preparation, Sequencing, Purification, SDS Page, Staining, Western Blot

    Characterization of cytophilic antibody responses. IgG1 ( a ) and IgG3 ( b ) antibody concentrations to EBA175RIII–V Ll in asymptomatic children from Obom and Abura measured in the peak malaria season (July). Processes similar to that described in Fig. 3 were used to determine the concentrations and avidity of IgG1 and IgG3, the only difference was that goat anti-human IgG1 and goat anti-human IgG3 secondary antibodies were used in place of the goat anti-human IgG. The graphs represent the median antibody concentrations with interquartile range as error bars

    Journal: Malaria Journal

    Article Title: Assessment of the quality and quantity of naturally induced antibody responses to EBA175RIII–V in Ghanaian children living in two communities with varying malaria transmission patterns

    doi: 10.1186/s12936-017-2167-3

    Figure Lengend Snippet: Characterization of cytophilic antibody responses. IgG1 ( a ) and IgG3 ( b ) antibody concentrations to EBA175RIII–V Ll in asymptomatic children from Obom and Abura measured in the peak malaria season (July). Processes similar to that described in Fig. 3 were used to determine the concentrations and avidity of IgG1 and IgG3, the only difference was that goat anti-human IgG1 and goat anti-human IgG3 secondary antibodies were used in place of the goat anti-human IgG. The graphs represent the median antibody concentrations with interquartile range as error bars

    Article Snippet: The western blot was probed with penta-His mouse IgG1 monoclonal antibodies (Thermo Scientific, USA) followed by alkaline phosphatase conjugated goat anti mouse IgG (H + L) secondary antibodies (Thermo Scientific, USA).

    Techniques:

    Characterization of IgG responses against EBA175RIII–V Ll . Antibody concentrations (ng/ml) of plasma obtained from the enrolled children from Obom and Abura ( a ) diluted 1:200 was determined using indirect ELISA and a EBA175RIII–V Ll as the antigen coated onto the ELISA plate and goat anti-human IgG used as the secondary antibody. The relative avidities of these same plasma samples were determined using a modified indirect ELISA assay where an incubation of the bound plasma samples obtained from children Obom and Abura ( b ) were treated with sodium thiocyanide is incorporated into the protocol. Plasma samples were obtained from whole blood collected from the children during the months of July 2015, October 2015 and February 2016. Data in the graphs are represented as the median with the interquartile range

    Journal: Malaria Journal

    Article Title: Assessment of the quality and quantity of naturally induced antibody responses to EBA175RIII–V in Ghanaian children living in two communities with varying malaria transmission patterns

    doi: 10.1186/s12936-017-2167-3

    Figure Lengend Snippet: Characterization of IgG responses against EBA175RIII–V Ll . Antibody concentrations (ng/ml) of plasma obtained from the enrolled children from Obom and Abura ( a ) diluted 1:200 was determined using indirect ELISA and a EBA175RIII–V Ll as the antigen coated onto the ELISA plate and goat anti-human IgG used as the secondary antibody. The relative avidities of these same plasma samples were determined using a modified indirect ELISA assay where an incubation of the bound plasma samples obtained from children Obom and Abura ( b ) were treated with sodium thiocyanide is incorporated into the protocol. Plasma samples were obtained from whole blood collected from the children during the months of July 2015, October 2015 and February 2016. Data in the graphs are represented as the median with the interquartile range

    Article Snippet: The western blot was probed with penta-His mouse IgG1 monoclonal antibodies (Thermo Scientific, USA) followed by alkaline phosphatase conjugated goat anti mouse IgG (H + L) secondary antibodies (Thermo Scientific, USA).

    Techniques: Indirect ELISA, Enzyme-linked Immunosorbent Assay, Modification, Incubation