goat anti mouse igg1  (SouthernBiotech)

 
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  • 99
    Name:
    Goat Anti Mouse IgG Fc BIOT
    Description:

    Catalog Number:
    1033-08
    Price:
    None
    Source:
    Pooled antisera from goats hyperimmunized with mouse IgG
    Applications:
    Quality tested applications for relevant formats include -ELISA 1FLISAOther referenced applications for relevant formats include -Western Blot 2
    Format:
    BIOT (Biotin)
    Isotype:
    Goat IgG
    Buy from Supplier


    Structured Review

    SouthernBiotech goat anti mouse igg1
    Goat Anti Mouse IgG Fc BIOT

    https://www.bioz.com/result/goat anti mouse igg1/product/SouthernBiotech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg1 - by Bioz Stars, 2021-04
    99/100 stars

    Images

    1) Product Images from "AP-1 Transcription Factor JunD Confers Protection from Accelerated Nephrotoxic Nephritis and Control Podocyte-Specific Vegfa Expression"

    Article Title: AP-1 Transcription Factor JunD Confers Protection from Accelerated Nephrotoxic Nephritis and Control Podocyte-Specific Vegfa Expression

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2011.03.006

    Deposited glomerular IgG and humoral immune response assessment in WT and Jund -/- mice 10 days after accelerated NTN induction. Quantitative immunofluorescence for sheep IgG ( A ) and mouse IgG ( B ). Representative glomeruli showing immunofluorescence for sheep and mouse IgG 10 days after the induction of accelerated NTN ( C ). Serum levels of mouse total IgG specific for sheep IgG ( D ). Ten mice were used in each group. AFU indicates arbitrary fluorescence unit; AEU, arbitrary enzyme-linked immunosorbent assay unit; ns, nonsignificant.
    Figure Legend Snippet: Deposited glomerular IgG and humoral immune response assessment in WT and Jund -/- mice 10 days after accelerated NTN induction. Quantitative immunofluorescence for sheep IgG ( A ) and mouse IgG ( B ). Representative glomeruli showing immunofluorescence for sheep and mouse IgG 10 days after the induction of accelerated NTN ( C ). Serum levels of mouse total IgG specific for sheep IgG ( D ). Ten mice were used in each group. AFU indicates arbitrary fluorescence unit; AEU, arbitrary enzyme-linked immunosorbent assay unit; ns, nonsignificant.

    Techniques Used: Mouse Assay, Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay

    2) Product Images from "Dendritic Cells Expressing MyD88 Molecule Are Necessary and Sufficient for CpG-Mediated Inhibition of IgE Production In Vivo"

    Article Title: Dendritic Cells Expressing MyD88 Molecule Are Necessary and Sufficient for CpG-Mediated Inhibition of IgE Production In Vivo

    Journal: Cells

    doi: 10.3390/cells8101165

    CpG-induced inhibition of IgE and augmented IgG2a production is independent of type I or type II interferon receptors. 129 WT strain or mice lacking type I interferon receptor (IFNAR −/− ) or type II interferon receptor (IFNGR −/− ) on 129 background were submitted to the same protocol of OVA sensitization with alum and i.n. OVA challenge. Experiments were performed on day 22. Serum levels of (A) total IgE, (B) OVA-specific IgE or (C) OVA-specific IgG2c were measured by ELISA. OVA groups ( n = 5) and OVA+CpG groups ( n = 5). Values represent the mean ± SD and are representative of two independent experiments. One-way ANOVA: *** p
    Figure Legend Snippet: CpG-induced inhibition of IgE and augmented IgG2a production is independent of type I or type II interferon receptors. 129 WT strain or mice lacking type I interferon receptor (IFNAR −/− ) or type II interferon receptor (IFNGR −/− ) on 129 background were submitted to the same protocol of OVA sensitization with alum and i.n. OVA challenge. Experiments were performed on day 22. Serum levels of (A) total IgE, (B) OVA-specific IgE or (C) OVA-specific IgG2c were measured by ELISA. OVA groups ( n = 5) and OVA+CpG groups ( n = 5). Values represent the mean ± SD and are representative of two independent experiments. One-way ANOVA: *** p

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

    CpG inhibits IgE and enhances IgG production. C57BL/6 wild-type (WT) mice were subcutaneously sensitized with ovalbumin (OVA) or OVA plus CpG (OVA+CpG) with or without alum adjuvant on days 0 and 7 and challenged intranasally with OVA on days 14 and 21. Experiments were performed on day 22. Control mice consisted of non-manipulated animals. ( A ) Schematic experimental protocols. Numbers of ( B ) Total Cells and ( C ) Eosinophils in BAL. Serum levels of ( D ) Total IgE, ( E ) OVA-specific IgE, ( F ) OVA-specific IgG2c and ( G ) OVA-specific IgG1. Groups sensitized with alum adjuvant ( n = 5) and without alum ( n = 3). Values represent the mean ± SD and are representative of two independent experiments. One-way ANOVA: ** p
    Figure Legend Snippet: CpG inhibits IgE and enhances IgG production. C57BL/6 wild-type (WT) mice were subcutaneously sensitized with ovalbumin (OVA) or OVA plus CpG (OVA+CpG) with or without alum adjuvant on days 0 and 7 and challenged intranasally with OVA on days 14 and 21. Experiments were performed on day 22. Control mice consisted of non-manipulated animals. ( A ) Schematic experimental protocols. Numbers of ( B ) Total Cells and ( C ) Eosinophils in BAL. Serum levels of ( D ) Total IgE, ( E ) OVA-specific IgE, ( F ) OVA-specific IgG2c and ( G ) OVA-specific IgG1. Groups sensitized with alum adjuvant ( n = 5) and without alum ( n = 3). Values represent the mean ± SD and are representative of two independent experiments. One-way ANOVA: ** p

    Techniques Used: Mouse Assay

    3) Product Images from "IgG4 Autoantibodies Attenuate Systemic Lupus Erythematosus Progression by Suppressing Complement Consumption and Inflammatory Cytokine Production"

    Article Title: IgG4 Autoantibodies Attenuate Systemic Lupus Erythematosus Progression by Suppressing Complement Consumption and Inflammatory Cytokine Production

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.01047

    Effects of IgG1 autoantibodies on renal function and renal pathology in MRL- lpr/lpr mice. Representative fluorescence microscopy images (A) and statistical analysis (B) of C1q, C3, IgG1, and IgG3 deposition in the kidney of non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice ( n = 6) at 19 weeks of age (scale bars, 50 μm). Serum creatinine, BUN, and urinary proteins (C) , and light-microscopy examination of the histopathology of glomerulonephritis using H E, PAS, and Masson's staining (D) in non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice ( n = 6) at 19 weeks of age (scale bars, 50 μm). * P
    Figure Legend Snippet: Effects of IgG1 autoantibodies on renal function and renal pathology in MRL- lpr/lpr mice. Representative fluorescence microscopy images (A) and statistical analysis (B) of C1q, C3, IgG1, and IgG3 deposition in the kidney of non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice ( n = 6) at 19 weeks of age (scale bars, 50 μm). Serum creatinine, BUN, and urinary proteins (C) , and light-microscopy examination of the histopathology of glomerulonephritis using H E, PAS, and Masson's staining (D) in non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice ( n = 6) at 19 weeks of age (scale bars, 50 μm). * P

    Techniques Used: Mouse Assay, Fluorescence, Microscopy, Light Microscopy, Histopathology, Staining

    Analysis of purified human IgG4 and the effect of IgG4 on complement consumption by autoantibody-autoantigen ICs in vitro . Human IgG4 from the sera of healthy control subjects ( n = 12) and newly diagnosed SLE patients ( n = 12) was purified by immunoaffinity column chromatography and the antibody mixtures were analyzed by SDS-PAGE (A) , IF staining with HEp-2 cells (scale bars, 50 μm) (B) , and the ANA Euroline Profile 3 Kit (C) . (D) Schematic representation of the in vitro complement-consumption assay. Three independent experiments were performed. Consumption of complement proteins including C3 (E) , C4 (F) , complement factors B (G) and H (H) , as well as C5b-9 deposits (I) in cultured HEp-2 cells were detected after co-culturing HEp-2 cells with SLE serum, mixed with or without different concentrations of purified IgG4 (the IgG4: IgG ratio was 50 or 100%). As blank control, the first column of white circles from the left represents only sera from SLE patients ( n = 12), without IgG4 and HEp-2 cells (E–I) ; As positive control of the existence of autoantigens, the second column of white circles represents sera from SLE patients ( n = 12), plus HEp-2 cells, but without IgG4 (E–I) . * P
    Figure Legend Snippet: Analysis of purified human IgG4 and the effect of IgG4 on complement consumption by autoantibody-autoantigen ICs in vitro . Human IgG4 from the sera of healthy control subjects ( n = 12) and newly diagnosed SLE patients ( n = 12) was purified by immunoaffinity column chromatography and the antibody mixtures were analyzed by SDS-PAGE (A) , IF staining with HEp-2 cells (scale bars, 50 μm) (B) , and the ANA Euroline Profile 3 Kit (C) . (D) Schematic representation of the in vitro complement-consumption assay. Three independent experiments were performed. Consumption of complement proteins including C3 (E) , C4 (F) , complement factors B (G) and H (H) , as well as C5b-9 deposits (I) in cultured HEp-2 cells were detected after co-culturing HEp-2 cells with SLE serum, mixed with or without different concentrations of purified IgG4 (the IgG4: IgG ratio was 50 or 100%). As blank control, the first column of white circles from the left represents only sera from SLE patients ( n = 12), without IgG4 and HEp-2 cells (E–I) ; As positive control of the existence of autoantigens, the second column of white circles represents sera from SLE patients ( n = 12), plus HEp-2 cells, but without IgG4 (E–I) . * P

    Techniques Used: Purification, In Vitro, Column Chromatography, SDS Page, Staining, Cell Culture, Positive Control

    Analysis of purified IgG4 autoantigens and their effect on inflammatory cytokine production by SLE PBMCs in vitro . Purified nuclear autoantigens from HEp-2 cells were analyzed by SDS-PAGE (A) , and serum anti-nuclear IgG autoantibodies from healthy control subjects and SLE patients were detected by ELISA (B) . (C) Schematic representation of the in vitro inflammatory cytokine assays. Three independent experiments were performed. The production of inflammatory cytokines, including IFN-γ (D) , IL-6 (E) , and IL-17 (F) by autoantigen-stimulated SLE PBMCs were detected, mixed with or without different concentrations of purified IgG4 (the IgG4: IgG ratio was 50 or 100%). As blank control, the first column of white circles from the left represents only SLE PBMCs ( n = 12, grown in DMEM/F12 supplemented with 25% serum of SLE patients), without IgG4 and nuclear antigens (D–F) ; As positive control of the existence of nuclear antigens, the second column of white circles represents SLE PBMCs ( n = 12, grown in DMEM/F12 supplemented with 25% serum of SLE patients), plus nuclear antigens, but without IgG4 (D–F) . * P
    Figure Legend Snippet: Analysis of purified IgG4 autoantigens and their effect on inflammatory cytokine production by SLE PBMCs in vitro . Purified nuclear autoantigens from HEp-2 cells were analyzed by SDS-PAGE (A) , and serum anti-nuclear IgG autoantibodies from healthy control subjects and SLE patients were detected by ELISA (B) . (C) Schematic representation of the in vitro inflammatory cytokine assays. Three independent experiments were performed. The production of inflammatory cytokines, including IFN-γ (D) , IL-6 (E) , and IL-17 (F) by autoantigen-stimulated SLE PBMCs were detected, mixed with or without different concentrations of purified IgG4 (the IgG4: IgG ratio was 50 or 100%). As blank control, the first column of white circles from the left represents only SLE PBMCs ( n = 12, grown in DMEM/F12 supplemented with 25% serum of SLE patients), without IgG4 and nuclear antigens (D–F) ; As positive control of the existence of nuclear antigens, the second column of white circles represents SLE PBMCs ( n = 12, grown in DMEM/F12 supplemented with 25% serum of SLE patients), plus nuclear antigens, but without IgG4 (D–F) . * P

    Techniques Used: Purification, In Vitro, SDS Page, Enzyme-linked Immunosorbent Assay, Positive Control

    Effect of IgG1 autoantibodies on complement consumption and inflammatory cytokine production in MRL- lpr/lpr mice. (A) Experimental design and timeline. Serum mouse IgG1 from control mice ( n = 20) and aged MRL- lpr/lpr mice ( n = 20) was purified by immunoaffinity-column chromatography and the antibody mixtures were analyzed by SDS-PAGE (B) and IF staining with HEp-2 cells (scale bars, 50 μm) (C) . (D,E) Serum levels of autoantibodies in non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice at 7, 11, 15, and 19 weeks of age ( n = 6). (F) Representative images of the spleens of non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice at 19 weeks of age (from left to right) and statistical analysis of the spleen weight as a percentage of the body weight ( n = 6). (G) Serum levels of complement proteins including C3, complement factor B, and complement factor H, and (H) inflammatory cytokines including IFN-γ, IL-6, and IL-17 in non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice at 19 weeks of age ( n = 6). * P
    Figure Legend Snippet: Effect of IgG1 autoantibodies on complement consumption and inflammatory cytokine production in MRL- lpr/lpr mice. (A) Experimental design and timeline. Serum mouse IgG1 from control mice ( n = 20) and aged MRL- lpr/lpr mice ( n = 20) was purified by immunoaffinity-column chromatography and the antibody mixtures were analyzed by SDS-PAGE (B) and IF staining with HEp-2 cells (scale bars, 50 μm) (C) . (D,E) Serum levels of autoantibodies in non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice at 7, 11, 15, and 19 weeks of age ( n = 6). (F) Representative images of the spleens of non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice at 19 weeks of age (from left to right) and statistical analysis of the spleen weight as a percentage of the body weight ( n = 6). (G) Serum levels of complement proteins including C3, complement factor B, and complement factor H, and (H) inflammatory cytokines including IFN-γ, IL-6, and IL-17 in non-treated and control or lupus IgG1-treated MRL- lpr/lpr mice at 19 weeks of age ( n = 6). * P

    Techniques Used: Mouse Assay, Purification, Column Chromatography, SDS Page, Staining

    Schematic representation of IgG4 autoantibody-mediated attenuation of disease progression in SLE by suppressing complement consumption and inflammatory cytokine production. During SLE progression, the inflammatory response induced by each autoreactive IgG subtype depends on its ability to activate effector function. IgG4 autoantibodies may bind autoantigens completely with other subclasses of autoantibodies such as IgG1, IgG2, and IgG3, to form autoantibody-autoantigen ICs. The hinge region of IgG4 has a unique structure that determines its limited ability to activate effector function. Thus, compared with other subclasses of autoantibodies, IgG4 autoantibodies may attenuate disease progression in SLE by suppressing complement consumption and inflammatory cytokine production.
    Figure Legend Snippet: Schematic representation of IgG4 autoantibody-mediated attenuation of disease progression in SLE by suppressing complement consumption and inflammatory cytokine production. During SLE progression, the inflammatory response induced by each autoreactive IgG subtype depends on its ability to activate effector function. IgG4 autoantibodies may bind autoantigens completely with other subclasses of autoantibodies such as IgG1, IgG2, and IgG3, to form autoantibody-autoantigen ICs. The hinge region of IgG4 has a unique structure that determines its limited ability to activate effector function. Thus, compared with other subclasses of autoantibodies, IgG4 autoantibodies may attenuate disease progression in SLE by suppressing complement consumption and inflammatory cytokine production.

    Techniques Used:

    Correlation between serum levels of IgG4 autoantibody and disease activity in patients with SLE. (A) Serum levels of ANA-IgG ( n = 106) and ANA-IgG4 in newly diagnosed patients in group I ( n = 37) and group II ( n = 32) were detected by IF staining of HEp-2 cells, and the fluorescence intensities were graded (scale bars, 50 μm). The SLEDAI score (B) , serum levels of inflammatory cytokines including IFN-γ, IL-6, and IL-17 (C) , serum levels of complement proteins including C3, C4, complement factor B, and complement factor H (D) , and serum creatinine, BUN, urine protein: creatinine, and 24-h proteinuria levels (E) in newly diagnosed patients with SLE of group I ( n = 37) and group II ( n = 32) were analyzed. * P
    Figure Legend Snippet: Correlation between serum levels of IgG4 autoantibody and disease activity in patients with SLE. (A) Serum levels of ANA-IgG ( n = 106) and ANA-IgG4 in newly diagnosed patients in group I ( n = 37) and group II ( n = 32) were detected by IF staining of HEp-2 cells, and the fluorescence intensities were graded (scale bars, 50 μm). The SLEDAI score (B) , serum levels of inflammatory cytokines including IFN-γ, IL-6, and IL-17 (C) , serum levels of complement proteins including C3, C4, complement factor B, and complement factor H (D) , and serum creatinine, BUN, urine protein: creatinine, and 24-h proteinuria levels (E) in newly diagnosed patients with SLE of group I ( n = 37) and group II ( n = 32) were analyzed. * P

    Techniques Used: Activity Assay, Staining, Fluorescence

    4) Product Images from "Effects of Repeated Chlamydia pneumoniae Inoculations on Aortic Lipid Accumulation and Inflammatory Response in C57BL/6J Mice †"

    Article Title: Effects of Repeated Chlamydia pneumoniae Inoculations on Aortic Lipid Accumulation and Inflammatory Response in C57BL/6J Mice †

    Journal:

    doi: 10.1128/IAI.73.10.6458-6466.2005

    Spearman's correlation between chlamydial Hsp60 IgG2c and mouse Hsp60 IgG levels in all mice (A) and in mice at 32 weeks of age (B).
    Figure Legend Snippet: Spearman's correlation between chlamydial Hsp60 IgG2c and mouse Hsp60 IgG levels in all mice (A) and in mice at 32 weeks of age (B).

    Techniques Used: Mouse Assay

    Total C. pneumoniae IgG antibody levels measured by the MIF and EIA methods and levels of mouse Hsp60 IgG antibodies measured by EIA. The study group, the number of inoculations given (e.g., six inoculations [6x]), and the age at which the first inoculation
    Figure Legend Snippet: Total C. pneumoniae IgG antibody levels measured by the MIF and EIA methods and levels of mouse Hsp60 IgG antibodies measured by EIA. The study group, the number of inoculations given (e.g., six inoculations [6x]), and the age at which the first inoculation

    Techniques Used: Enzyme-linked Immunosorbent Assay

    5) Product Images from "CpG Oligodeoxynucleotides Act as Adjuvants that Switch on T Helper 1 (Th1) Immunity "

    Article Title: CpG Oligodeoxynucleotides Act as Adjuvants that Switch on T Helper 1 (Th1) Immunity

    Journal: The Journal of Experimental Medicine

    doi:

    Th1-associated antigen-specific IgG2a responses are induced by immunization of BALB/c mice with IFA-HEL-CpG ODN but not IFA-HEL-non–CpG ODN. ( A–C ). Mice were injected i.p. with CFA-HEL (a control for a Th1-dominated response), IFA-HEL (a control for a Th2-dominated response), or IFA-HEL with 100 μg of CpG ODN 1826 or non–CpG ODN 1745. Sera were collected from mice 15–18 d after injection and assayed by ELISA for: ( A ) anti-HEL IgG2a, an isotype associated with Th1-dominated responses; ( B ) anti-HEL IgG1; and ( C ) anti-HEL total Ig response. A–C represent data from a single experiment representative of three similar experiments. ( D ) BALB/c mice were immunized as above, except that 30 μg of CpG ODN 1585, non–CpG ODN 1972, CpG ODN 1760, or non–CpG ODN 1908 was used for each mouse. Anti-HEL IgG2a antibodies were detected by serum ELISA. Data shown in D are representative of three similar experiments.
    Figure Legend Snippet: Th1-associated antigen-specific IgG2a responses are induced by immunization of BALB/c mice with IFA-HEL-CpG ODN but not IFA-HEL-non–CpG ODN. ( A–C ). Mice were injected i.p. with CFA-HEL (a control for a Th1-dominated response), IFA-HEL (a control for a Th2-dominated response), or IFA-HEL with 100 μg of CpG ODN 1826 or non–CpG ODN 1745. Sera were collected from mice 15–18 d after injection and assayed by ELISA for: ( A ) anti-HEL IgG2a, an isotype associated with Th1-dominated responses; ( B ) anti-HEL IgG1; and ( C ) anti-HEL total Ig response. A–C represent data from a single experiment representative of three similar experiments. ( D ) BALB/c mice were immunized as above, except that 30 μg of CpG ODN 1585, non–CpG ODN 1972, CpG ODN 1760, or non–CpG ODN 1908 was used for each mouse. Anti-HEL IgG2a antibodies were detected by serum ELISA. Data shown in D are representative of three similar experiments.

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

    6) Product Images from "One-step generation of monoclonal B cell receptor mice capable of isotype switching and somatic hypermutation"

    Article Title: One-step generation of monoclonal B cell receptor mice capable of isotype switching and somatic hypermutation

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20172064

    Isotype switching and plasmablast differentiation in Igh CGG/+ mice in vitro. (a) In vitro isotype switching. Purified Igh CGG/+ (CD45.2/2) and WT (CD45.1/1) B cells were cocultured in vitro with LPS and IL-4 to induce isotype switching to IgG 1 . Histogram shows IgG 1 staining by flow cytometry on day 3 of culture. Data are from three independent experiments and are quantified in the graph; proportions of IgG 1 + cells in Igh CGG/+ and WT B cells from the same experiment are connected by a line. P = 0.74 (unpaired t test). (b) AFC cell formation (as defined by CD138 expression) and IgG 1 secretion in purified B cells stimulated with LPS and IL-4 in vitro. Cells were presorted as in Fig. 3 a , and non–CGG-binding B cells were excluded from Igh CGG/+ cultures. Histograms are representative of, and graphs show pooled data for two independent experiments. n = 3 mice for WT and n = 6 mice for Igh CGG/+ CGG binding B cells. P = 0.33 for CD138 expression and P = 0.37 for supernatant IgG 1 . Numbers within flow plots indicate percentage of cells in the designated gate.
    Figure Legend Snippet: Isotype switching and plasmablast differentiation in Igh CGG/+ mice in vitro. (a) In vitro isotype switching. Purified Igh CGG/+ (CD45.2/2) and WT (CD45.1/1) B cells were cocultured in vitro with LPS and IL-4 to induce isotype switching to IgG 1 . Histogram shows IgG 1 staining by flow cytometry on day 3 of culture. Data are from three independent experiments and are quantified in the graph; proportions of IgG 1 + cells in Igh CGG/+ and WT B cells from the same experiment are connected by a line. P = 0.74 (unpaired t test). (b) AFC cell formation (as defined by CD138 expression) and IgG 1 secretion in purified B cells stimulated with LPS and IL-4 in vitro. Cells were presorted as in Fig. 3 a , and non–CGG-binding B cells were excluded from Igh CGG/+ cultures. Histograms are representative of, and graphs show pooled data for two independent experiments. n = 3 mice for WT and n = 6 mice for Igh CGG/+ CGG binding B cells. P = 0.33 for CD138 expression and P = 0.37 for supernatant IgG 1 . Numbers within flow plots indicate percentage of cells in the designated gate.

    Techniques Used: Mouse Assay, In Vitro, Purification, Staining, Flow Cytometry, Cytometry, Expressing, Binding Assay

    CSR, SHM, and antigen-driven selection in Igh CGG/+ B cells. (a) Igh CGG/+ B cells (CD45.2/2) were adoptively transferred into WT (CD45.1/1) recipients, which were immunized 1 d later with CGG in alum. CSR to IgG 1 was determined in GC B cells (CD19 + TCRβ – Fas + CD38 lo ) from donor (CD45.2/2) and recipient (CD45.1/1) mice by flow cytometry. Graph shows percentage of IgG 1 + GC B cells in five mice from two independent experiments. Proportions of switched cells in the same mouse are connected by a line. P = 0.23 (unpaired t test). Numbers within flow plots indicate percentage of cells in the designated gate. (b) Left: Serum titer of IgG 1 + /human Cκ + Ig in three mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG. Graph shows data for three mice from two independent experiments. Right: Western blot for human Igκ in serum of mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG 14 d after immunization. NR, nonreducing conditions; R, reducing conditions (DTT). Blot is representative of two experiments. (c–f) GC B cells from adoptive transfer experiments as in panel a were single sorted, and the entire Igh CGG/+ allele was PCR-amplified and sequenced. (c) Nucleotide mutation frequencies along the Igh CGG/+ locus, calculated as the fraction of times a particular nucleotide was mutated from the original sequence. (d) Total mutation frequency per region (normalized to sequence length). Each symbol represents one cell. (e) Number of nucleotide mutations in VDJ H and VJκ in Igh CGG/+ GC B cells compared with WT GC B cells carrying the same rearrangement selected by CGG immunization, as reported previously ( Tas et al., 2016 ; only data from LN#2 GC2 in Fig. 4 from Tas et al., 2016 are analyzed). Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. **, P ≤ 0.001; ****, P ≤ 0.0001 (unpaired t test). (f) Comparison of nucleotide mutation patterns between Igh CGG/+ B cells and WT GC B cells carrying the same rearrangement. Mutation patterns are shown for FR1 to FR4 regions of IgH and CDR1 to FR4 regions of Igκ. Shared nucleotide mutation positions are shown in red. Asterisk indicates a C119 > G mutation, which confers an approximately eightfold gain in affinity. Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. (g) Reconstructed phylogeny of Igh CGG/+ GC B cell VJ Igκ sequences from multiple GCs in one LN. Clades of expanded clones are indicated in red, green, and blue. Cells containing the high-affinity C119 > G mutation are indicated in green.
    Figure Legend Snippet: CSR, SHM, and antigen-driven selection in Igh CGG/+ B cells. (a) Igh CGG/+ B cells (CD45.2/2) were adoptively transferred into WT (CD45.1/1) recipients, which were immunized 1 d later with CGG in alum. CSR to IgG 1 was determined in GC B cells (CD19 + TCRβ – Fas + CD38 lo ) from donor (CD45.2/2) and recipient (CD45.1/1) mice by flow cytometry. Graph shows percentage of IgG 1 + GC B cells in five mice from two independent experiments. Proportions of switched cells in the same mouse are connected by a line. P = 0.23 (unpaired t test). Numbers within flow plots indicate percentage of cells in the designated gate. (b) Left: Serum titer of IgG 1 + /human Cκ + Ig in three mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG. Graph shows data for three mice from two independent experiments. Right: Western blot for human Igκ in serum of mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG 14 d after immunization. NR, nonreducing conditions; R, reducing conditions (DTT). Blot is representative of two experiments. (c–f) GC B cells from adoptive transfer experiments as in panel a were single sorted, and the entire Igh CGG/+ allele was PCR-amplified and sequenced. (c) Nucleotide mutation frequencies along the Igh CGG/+ locus, calculated as the fraction of times a particular nucleotide was mutated from the original sequence. (d) Total mutation frequency per region (normalized to sequence length). Each symbol represents one cell. (e) Number of nucleotide mutations in VDJ H and VJκ in Igh CGG/+ GC B cells compared with WT GC B cells carrying the same rearrangement selected by CGG immunization, as reported previously ( Tas et al., 2016 ; only data from LN#2 GC2 in Fig. 4 from Tas et al., 2016 are analyzed). Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. **, P ≤ 0.001; ****, P ≤ 0.0001 (unpaired t test). (f) Comparison of nucleotide mutation patterns between Igh CGG/+ B cells and WT GC B cells carrying the same rearrangement. Mutation patterns are shown for FR1 to FR4 regions of IgH and CDR1 to FR4 regions of Igκ. Shared nucleotide mutation positions are shown in red. Asterisk indicates a C119 > G mutation, which confers an approximately eightfold gain in affinity. Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. (g) Reconstructed phylogeny of Igh CGG/+ GC B cell VJ Igκ sequences from multiple GCs in one LN. Clades of expanded clones are indicated in red, green, and blue. Cells containing the high-affinity C119 > G mutation are indicated in green.

    Techniques Used: Selection, Mouse Assay, Flow Cytometry, Cytometry, Western Blot, Adoptive Transfer Assay, Polymerase Chain Reaction, Amplification, Mutagenesis, Sequencing, Clone Assay

    7) Product Images from "Identification of T helper (Th)1- and Th2-associated antigens of Cryptococcus neoformans in a murine model of pulmonary infection"

    Article Title: Identification of T helper (Th)1- and Th2-associated antigens of Cryptococcus neoformans in a murine model of pulmonary infection

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-21039-z

    Total and C. neoformans -specific levels of Th2-dependent IgG1 and IgE predominate in sera from susceptible wild-type (WT) and IL-12-deficient mice with pulmonary cryptococcosis. Total IgG1 ( a ) and IgE ( b ) levels increased significantly after infection with C. neoformans for most genotypes, whereas IgG2a levels ( c ) were not influenced by C. neoformans . Compared to wild-type mice, IL-4Rα-deficient mice had markedly lower levels of immunoglobulin (Ig)G1 ( a ), no production of IgE ( b ) and similar levels of total IgG2a ( c ). Opposite, IL-12-deficient mice showed similar IgG1 and significantly higher IgE levels ( a , b ), while reduced levels of IgG2a ( c ), compared to wild-type mice. Titers of C. neoformans -specific IgG2a ( d ) and IgG1 ( e ) antibodies in infected (infec.) mice reflected the distribution observed for total immunoglobulin levels. C. neoformans -specific IgG1 and IgG2a antibodies were absent in sera of naïve mice. A strong correlation was observed between total and C. neoformans -specific IgG1 levels ( f ) but not between total and C. neoformans -specific IgG2a levels ( g ) in infected mice of all genotypes. Each spot represents a serum sample of an individual mouse (2 to 14 animals per group from at least two independent experiments) with the line indicating the median. Serum samples were obtained from mice intranasally infected with 500 colony forming units of C. neoformans strain 1841 after about 56 days post infection. Statistical significance determined by Mann-Whitney U test is shown as following: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. Correlation was determined by nonparametric Spearman’s correlation test.
    Figure Legend Snippet: Total and C. neoformans -specific levels of Th2-dependent IgG1 and IgE predominate in sera from susceptible wild-type (WT) and IL-12-deficient mice with pulmonary cryptococcosis. Total IgG1 ( a ) and IgE ( b ) levels increased significantly after infection with C. neoformans for most genotypes, whereas IgG2a levels ( c ) were not influenced by C. neoformans . Compared to wild-type mice, IL-4Rα-deficient mice had markedly lower levels of immunoglobulin (Ig)G1 ( a ), no production of IgE ( b ) and similar levels of total IgG2a ( c ). Opposite, IL-12-deficient mice showed similar IgG1 and significantly higher IgE levels ( a , b ), while reduced levels of IgG2a ( c ), compared to wild-type mice. Titers of C. neoformans -specific IgG2a ( d ) and IgG1 ( e ) antibodies in infected (infec.) mice reflected the distribution observed for total immunoglobulin levels. C. neoformans -specific IgG1 and IgG2a antibodies were absent in sera of naïve mice. A strong correlation was observed between total and C. neoformans -specific IgG1 levels ( f ) but not between total and C. neoformans -specific IgG2a levels ( g ) in infected mice of all genotypes. Each spot represents a serum sample of an individual mouse (2 to 14 animals per group from at least two independent experiments) with the line indicating the median. Serum samples were obtained from mice intranasally infected with 500 colony forming units of C. neoformans strain 1841 after about 56 days post infection. Statistical significance determined by Mann-Whitney U test is shown as following: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. Correlation was determined by nonparametric Spearman’s correlation test.

    Techniques Used: Mouse Assay, Infection, MANN-WHITNEY

    8) Product Images from "Vaccination with Venezuelan Equine Encephalitis Replicons Encoding Cowpox Virus Structural Proteins Protects Mice from Intranasal Cowpox Virus Challenge"

    Article Title: Vaccination with Venezuelan Equine Encephalitis Replicons Encoding Cowpox Virus Structural Proteins Protects Mice from Intranasal Cowpox Virus Challenge

    Journal: Virology

    doi: 10.1016/j.virol.2007.01.001

    B5, A33, and A27-VRP elicit an IgG2a polarized antibody response
    Figure Legend Snippet: B5, A33, and A27-VRP elicit an IgG2a polarized antibody response

    Techniques Used:

    B5-VRP induces long-lasting, high levels of anti-B5 serum IgG
    Figure Legend Snippet: B5-VRP induces long-lasting, high levels of anti-B5 serum IgG

    Techniques Used:

    9) Product Images from "Resistance to visceral leishmaniasis is severely compromised in mice deficient of bradykinin B2-receptors"

    Article Title: Resistance to visceral leishmaniasis is severely compromised in mice deficient of bradykinin B2-receptors

    Journal: Parasites & Vectors

    doi: 10.1186/1756-3305-5-261

    Correlation between the clinical signs of visceral leishmaniasis and the cellular versus antibody responses to leishmanial antigen. Data represented in bivariate graphics: ( A ) the positive correlation between the individual increase of spleen and liver relative weights and the increase in liver parasite load; and ( B ) the negative correlation between the increase in liver relative weight and the decrease in DTH response to leishmanial antigen, and ( C ) the increase in liver relative weight and the decrease in anti-NH36 IgG2b antibody response. The results were expressed on graphs as scattering of individual values. R-squared (R 2 ) (coefficients of determination) estimates are shown on graphs. Trend lines were added.
    Figure Legend Snippet: Correlation between the clinical signs of visceral leishmaniasis and the cellular versus antibody responses to leishmanial antigen. Data represented in bivariate graphics: ( A ) the positive correlation between the individual increase of spleen and liver relative weights and the increase in liver parasite load; and ( B ) the negative correlation between the increase in liver relative weight and the decrease in DTH response to leishmanial antigen, and ( C ) the increase in liver relative weight and the decrease in anti-NH36 IgG2b antibody response. The results were expressed on graphs as scattering of individual values. R-squared (R 2 ) (coefficients of determination) estimates are shown on graphs. Trend lines were added.

    Techniques Used:

    Evaluation of the IgG1 and IgG2b response in sera of mice deficient of B 2 R. ELISA assay showing anti-Nucleoside hydrolase (NH36) and anti- L.(L.) chagasi promastigote lysate IgG1 and IgG2b antibodies in B 2 R +/+ wild type (C57) and B 2 R −/− mice (KOB2) intravenously infected with 3 x 10 7 amastigotes of Leishmania. (L.) chagasi. Results are shown as the individual absorbency values of 1/100 diluted sera obtained from two (NH36) or one (lysate) experiment with 5 mice per treatment in each experiment, at day 30 after infection. Asterisks indicate significant differences between groups as disclosed by the Mann Whitney analysis.
    Figure Legend Snippet: Evaluation of the IgG1 and IgG2b response in sera of mice deficient of B 2 R. ELISA assay showing anti-Nucleoside hydrolase (NH36) and anti- L.(L.) chagasi promastigote lysate IgG1 and IgG2b antibodies in B 2 R +/+ wild type (C57) and B 2 R −/− mice (KOB2) intravenously infected with 3 x 10 7 amastigotes of Leishmania. (L.) chagasi. Results are shown as the individual absorbency values of 1/100 diluted sera obtained from two (NH36) or one (lysate) experiment with 5 mice per treatment in each experiment, at day 30 after infection. Asterisks indicate significant differences between groups as disclosed by the Mann Whitney analysis.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Infection, MANN-WHITNEY

    10) Product Images from "The generation and evaluation of two panels of epitope-matched mouse IgG1, IgG2a, IgG2b and IgG3 antibodies specific for Plasmodium falciparum and Plasmodium yoelii merozoite surface protein 1-19 (MSP119)"

    Article Title: The generation and evaluation of two panels of epitope-matched mouse IgG1, IgG2a, IgG2b and IgG3 antibodies specific for Plasmodium falciparum and Plasmodium yoelii merozoite surface protein 1-19 (MSP119)

    Journal: Experimental Parasitology

    doi: 10.1016/j.exppara.2012.02.003

    Characterization of purified mouse IgG1-C1 by immunofluorescence assay (IFA). Mouse IgG1 is reactive with MSP1 19 on methanol–acetone-fixed smears of merozoites and erythrocytes infected with rodent P. berghei Tg for P. falciparum MSP1 19 (Pb-PfM19). Top panel: mAbs 12.8 and 12.10 previously described and known not to bind to Pb-PfM19 ( Lazarou et al., 2009 ) and B10 anti- Py MSP1 19 as negative control ( Spencer Valero et al., 1998 ). Bottom panel: Human IgG1-C1 (JS1) and human IgG1-e9 (JS2) as previously described ( McIntosh et al., 2007 ) and murine IgG1-C1 bind Pb-PfM19 infected erythrocytes. Binding was visualized by staining with a FITC-conjugated secondary reagent with nuclei stained with DAPI and assessed by fluorescent microscopy under 40× magnification.
    Figure Legend Snippet: Characterization of purified mouse IgG1-C1 by immunofluorescence assay (IFA). Mouse IgG1 is reactive with MSP1 19 on methanol–acetone-fixed smears of merozoites and erythrocytes infected with rodent P. berghei Tg for P. falciparum MSP1 19 (Pb-PfM19). Top panel: mAbs 12.8 and 12.10 previously described and known not to bind to Pb-PfM19 ( Lazarou et al., 2009 ) and B10 anti- Py MSP1 19 as negative control ( Spencer Valero et al., 1998 ). Bottom panel: Human IgG1-C1 (JS1) and human IgG1-e9 (JS2) as previously described ( McIntosh et al., 2007 ) and murine IgG1-C1 bind Pb-PfM19 infected erythrocytes. Binding was visualized by staining with a FITC-conjugated secondary reagent with nuclei stained with DAPI and assessed by fluorescent microscopy under 40× magnification.

    Techniques Used: Purification, Immunofluorescence, Infection, Negative Control, Binding Assay, Staining, Microscopy

    Mouse Pf MSP1 19 -specific IgG1 is functional. Neutrophil NADH oxidative bursts were measured by adding antibodies to a chemiluminescence plate coated with recombinant purified antigen Pf MSP1 19 . Relative Light Units (RLU, arbitrary units of light produced/second) were calculated from triplicate wells using neutrophils as previously described ( McIntosh et al., 2007; Pleass et al., 2007, 2003 ). Each point in the graph represents the mean value of triplicate wells. RLU were measured for over 120 min after the IgGs were added to neutrophils at two different concentrations of 50 (△) or 100 (○) μg/ml of mIgG1-C1. The overall response was comparable to that derived from human IgG1-C1 at the same concentrations (▴, ● 50 or 100 μg/ml respectively). An IgG2a-Fc fused to antigen ( Pf MSP1 19 -IgG2a-Fc) was used as an internal negative control ( , 50 or 100 μg/ml respectively).
    Figure Legend Snippet: Mouse Pf MSP1 19 -specific IgG1 is functional. Neutrophil NADH oxidative bursts were measured by adding antibodies to a chemiluminescence plate coated with recombinant purified antigen Pf MSP1 19 . Relative Light Units (RLU, arbitrary units of light produced/second) were calculated from triplicate wells using neutrophils as previously described ( McIntosh et al., 2007; Pleass et al., 2007, 2003 ). Each point in the graph represents the mean value of triplicate wells. RLU were measured for over 120 min after the IgGs were added to neutrophils at two different concentrations of 50 (△) or 100 (○) μg/ml of mIgG1-C1. The overall response was comparable to that derived from human IgG1-C1 at the same concentrations (▴, ● 50 or 100 μg/ml respectively). An IgG2a-Fc fused to antigen ( Pf MSP1 19 -IgG2a-Fc) was used as an internal negative control ( , 50 or 100 μg/ml respectively).

    Techniques Used: Functional Assay, Recombinant, Purification, Produced, Derivative Assay, Negative Control

    Surface plasmon resonance (SPR)-derived association and dissociation curves of mouse IgGs binding to Pf MSP1 19 . SPR association ( K A ) and dissociation ( K D ) curves of mouse IgG binding to recombinant purified antigen Pf MSP1 19 amine-coupled and immobilized to a CM5 sensor chip. The antibodies were injected into flow at five different concentrations of 2 μM (red), 1 μM (green), 0.5 μM (cyan), 0.25 μM pink), and 0.125 μM (dark blue) at time 0 and replaced with buffer alone at the times indicated with arrows. No binding was seen from any of the IgGs onto control Ag coated in flow cell 2. Data are summarized in Table 1 . (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
    Figure Legend Snippet: Surface plasmon resonance (SPR)-derived association and dissociation curves of mouse IgGs binding to Pf MSP1 19 . SPR association ( K A ) and dissociation ( K D ) curves of mouse IgG binding to recombinant purified antigen Pf MSP1 19 amine-coupled and immobilized to a CM5 sensor chip. The antibodies were injected into flow at five different concentrations of 2 μM (red), 1 μM (green), 0.5 μM (cyan), 0.25 μM pink), and 0.125 μM (dark blue) at time 0 and replaced with buffer alone at the times indicated with arrows. No binding was seen from any of the IgGs onto control Ag coated in flow cell 2. Data are summarized in Table 1 . (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Techniques Used: SPR Assay, Derivative Assay, Binding Assay, Recombinant, Purification, Chromatin Immunoprecipitation, Injection, Flow Cytometry

    Generating two panels of epitope-matched murine IgG Abs specific for MSP1 19 epitopes C1 ( P. falciparum - Pf MSP1 19 ) or B10 ( P. yoelii - Py MSP1 19 ). Diagram represents a general overview to the construction of (a) C1 and (b) B10 mouse IgGs. Variable genes from parental mAbs C1 or B10 were sub-cloned into redesigned expression vectors for each of the mouse IgG subclasses.
    Figure Legend Snippet: Generating two panels of epitope-matched murine IgG Abs specific for MSP1 19 epitopes C1 ( P. falciparum - Pf MSP1 19 ) or B10 ( P. yoelii - Py MSP1 19 ). Diagram represents a general overview to the construction of (a) C1 and (b) B10 mouse IgGs. Variable genes from parental mAbs C1 or B10 were sub-cloned into redesigned expression vectors for each of the mouse IgG subclasses.

    Techniques Used: Clone Assay, Expressing

    Sequencing of C1 and B10 VH and VL genes. Amino acid sequences alignment of Pf MSP1 19 and Py MSP1 19 -binding epitope-matched mouse IgGs with the respective parental expression plasmids. (a) pVL-C1-mouse kappa compared with pVKExpress C1. (b) IgG1, IgG2a, IgG2b, and IgG3-C1 mouse IgG constructs compared with (h)IgG1-C1. (c) pVL-B10-mouse kappa compared with pB10-VL/10. (d) IgG1, IgG2a, IgG2b, and IgG3-B10 mouse IgG constructs compared with pB10-VH/5.
    Figure Legend Snippet: Sequencing of C1 and B10 VH and VL genes. Amino acid sequences alignment of Pf MSP1 19 and Py MSP1 19 -binding epitope-matched mouse IgGs with the respective parental expression plasmids. (a) pVL-C1-mouse kappa compared with pVKExpress C1. (b) IgG1, IgG2a, IgG2b, and IgG3-C1 mouse IgG constructs compared with (h)IgG1-C1. (c) pVL-B10-mouse kappa compared with pB10-VL/10. (d) IgG1, IgG2a, IgG2b, and IgG3-B10 mouse IgG constructs compared with pB10-VH/5.

    Techniques Used: Sequencing, Binding Assay, Expressing, Construct

    Characterization of purified recombinant Pf MSP1 19 -specific epitope-matched mouse IgGs. Characterization of the purified Pf MSP1 19 -specific mouse IgGs. KEY: IgG1 (A), IgG2a, (B), IgG2b (C), IgG3 (D). (a) Five micrograms of each Ab was run on NuPAGE 4–12% Bis–Tris SDS gels (Invitrogen) under denaturing non-reducing conditions (A–D) or denaturing reducing conditions (E–H) and compared with the control antibody human IgG1 (JS1) from which they were derived (IDEM). An expected MW of ∼150 kDa for the intact antibodies was shown (arrowed, ‘IgG’) although free constitutive heavy chains and light chain were observed (50 and 25 kDa respectively, arrowed). (b) Western Blot analysis of the IgG samples transferred into PVDF membranes. Analysis was made following the same conditions and sample order described on (a). Murine IgG1, IgG2a-C1 antibodies were clearly detectable with polyclonal anti-mouse IgG Fc-specific secondary antibody compared to IgG2b which was detected very faintly, but none of them with the subclass-specific secondary antibodies used in ELISA.
    Figure Legend Snippet: Characterization of purified recombinant Pf MSP1 19 -specific epitope-matched mouse IgGs. Characterization of the purified Pf MSP1 19 -specific mouse IgGs. KEY: IgG1 (A), IgG2a, (B), IgG2b (C), IgG3 (D). (a) Five micrograms of each Ab was run on NuPAGE 4–12% Bis–Tris SDS gels (Invitrogen) under denaturing non-reducing conditions (A–D) or denaturing reducing conditions (E–H) and compared with the control antibody human IgG1 (JS1) from which they were derived (IDEM). An expected MW of ∼150 kDa for the intact antibodies was shown (arrowed, ‘IgG’) although free constitutive heavy chains and light chain were observed (50 and 25 kDa respectively, arrowed). (b) Western Blot analysis of the IgG samples transferred into PVDF membranes. Analysis was made following the same conditions and sample order described on (a). Murine IgG1, IgG2a-C1 antibodies were clearly detectable with polyclonal anti-mouse IgG Fc-specific secondary antibody compared to IgG2b which was detected very faintly, but none of them with the subclass-specific secondary antibodies used in ELISA.

    Techniques Used: Purification, Recombinant, Derivative Assay, Western Blot, Enzyme-linked Immunosorbent Assay

    Passive transfer of mouse Pf MSP1 19 -specific IgG1 does not protect mice from infection. Groups of four BALB/c wild type littermates were injected i.p. with a total dose of 1.5 mg of purified anti- P. falciparum mouse IgG1 (open symbols, groups 1 and 3) or with PBS-vehicle control (closed symbols, groups 2 and 4) and challenged with 5000 parasitized red blood cells (prbc) derived from ‘passage’ animals infected with Pb-PfM19. Similar results were obtained from two independent experiments (a–b and c–d respectively). (a) and (c) Mean weight as % of initial (mean weight from days −1, 0, +1) was assessed daily to determine if an animal had lost more than 20% of its original weight (dotted line) used as a humane end-point in accordance with Home Office regulations. (c) and (d) Seven out of eight animals treated with mIgG1-C1 succumbed to malaria infection and no significant difference was shown between the test and control groups. The percentage of parasitemia between the test and control groups was similar in both experiments.
    Figure Legend Snippet: Passive transfer of mouse Pf MSP1 19 -specific IgG1 does not protect mice from infection. Groups of four BALB/c wild type littermates were injected i.p. with a total dose of 1.5 mg of purified anti- P. falciparum mouse IgG1 (open symbols, groups 1 and 3) or with PBS-vehicle control (closed symbols, groups 2 and 4) and challenged with 5000 parasitized red blood cells (prbc) derived from ‘passage’ animals infected with Pb-PfM19. Similar results were obtained from two independent experiments (a–b and c–d respectively). (a) and (c) Mean weight as % of initial (mean weight from days −1, 0, +1) was assessed daily to determine if an animal had lost more than 20% of its original weight (dotted line) used as a humane end-point in accordance with Home Office regulations. (c) and (d) Seven out of eight animals treated with mIgG1-C1 succumbed to malaria infection and no significant difference was shown between the test and control groups. The percentage of parasitemia between the test and control groups was similar in both experiments.

    Techniques Used: Mouse Assay, Infection, Injection, Purification, Derivative Assay

    11) Product Images from "Disparate T cell requirements of two subsets of lupus-specific autoantibodies in pristane-treated mice"

    Article Title: Disparate T cell requirements of two subsets of lupus-specific autoantibodies in pristane-treated mice

    Journal: Clinical and Experimental Immunology

    doi: 10.1046/j.1365-2249.1999.00825.x

    Renal pathology. Top, frozen sections of renal tissue from PBS-treated BALB/c nu/nu mice were stained with FITC-conjugated goat anti-mouse IgG1. (A) Diffuse mesangial staining. (B) Mesangio-capillary distribution of IgG1. Note that relatively marked renal immune complex deposition may occur spontaneously in nude mice. Bottom: (A) Tissue from a PBS-treated BALB/c nu/nu mouse exhibiting numerous mesangial electron-dense deposits (arrow). (B) Tissue from a pristane-treated BALB/c nu/nu mouse exhibiting mesangial (straight arrow) and subepithelial (curved arrow) dense deposits.
    Figure Legend Snippet: Renal pathology. Top, frozen sections of renal tissue from PBS-treated BALB/c nu/nu mice were stained with FITC-conjugated goat anti-mouse IgG1. (A) Diffuse mesangial staining. (B) Mesangio-capillary distribution of IgG1. Note that relatively marked renal immune complex deposition may occur spontaneously in nude mice. Bottom: (A) Tissue from a PBS-treated BALB/c nu/nu mouse exhibiting numerous mesangial electron-dense deposits (arrow). (B) Tissue from a pristane-treated BALB/c nu/nu mouse exhibiting mesangial (straight arrow) and subepithelial (curved arrow) dense deposits.

    Techniques Used: Mouse Assay, Staining

    Anti-ssDNA antibodies in pristane-treated mice. Top, levels of IgM anti-ssDNA antibodies by ELISA in sera obtained 2 weeks after pristane or PBS treatment. Bottom, levels of IgG anti-ssDNA antibodies by ELISA in sera obtained 5 months after pristane or PBS treatment. Groups of mice are indicated below. All sera were tested at a 1:500 dilution.
    Figure Legend Snippet: Anti-ssDNA antibodies in pristane-treated mice. Top, levels of IgM anti-ssDNA antibodies by ELISA in sera obtained 2 weeks after pristane or PBS treatment. Bottom, levels of IgG anti-ssDNA antibodies by ELISA in sera obtained 5 months after pristane or PBS treatment. Groups of mice are indicated below. All sera were tested at a 1:500 dilution.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    12) Product Images from "Tropomyosin implicated in host protective responses to microfilariae in onchocerciasis"

    Article Title: Tropomyosin implicated in host protective responses to microfilariae in onchocerciasis

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

    doi:

    Comparison of the B cell epitope of O. volvulus tropomyosin with the equivalent region of tropomyosins from other organisms. The deduced amino acid sequence shown is that of the shortest peptide recognized by IgG in serum from mice vaccinated with a PBS extract of microfilariae. Aligned are representative sequences from nematodes, an insect, and mammals. The IgE-binding epitope of shrimp tropomyosin also is shown. Amino acids identical to those of the O. volvulus epitope are represented by a dot.
    Figure Legend Snippet: Comparison of the B cell epitope of O. volvulus tropomyosin with the equivalent region of tropomyosins from other organisms. The deduced amino acid sequence shown is that of the shortest peptide recognized by IgG in serum from mice vaccinated with a PBS extract of microfilariae. Aligned are representative sequences from nematodes, an insect, and mammals. The IgE-binding epitope of shrimp tropomyosin also is shown. Amino acids identical to those of the O. volvulus epitope are represented by a dot.

    Techniques Used: Sequencing, Mouse Assay, Binding Assay

    ELISA analysis of the IgG response to MOv14 of human onchocerciasis infection sera. ( a ) Comparison of the response of individuals from two onchocerciasis endemic foci with uninfected British controls. Group mean values are represented by a short, horizontal line. The cut-off value is represented by a horizontal line across the graph and is derived from the mean of the control values plus 2 SD. Relationship between microfilarial skin density and anti-MOv14 IgG levels in a Mali focus ( b ) and an Ecuador focus ( c ).
    Figure Legend Snippet: ELISA analysis of the IgG response to MOv14 of human onchocerciasis infection sera. ( a ) Comparison of the response of individuals from two onchocerciasis endemic foci with uninfected British controls. Group mean values are represented by a short, horizontal line. The cut-off value is represented by a horizontal line across the graph and is derived from the mean of the control values plus 2 SD. Relationship between microfilarial skin density and anti-MOv14 IgG levels in a Mali focus ( b ) and an Ecuador focus ( c ).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Derivative Assay

    13) Product Images from "Abated immune responses against recombinant hepatitis-b vaccine by chitin in mice"

    Article Title: Abated immune responses against recombinant hepatitis-b vaccine by chitin in mice

    Journal: bioRxiv

    doi: 10.1101/2021.03.25.436808

    Effect of HBsAg-Chitin formulation on IgG titre of Mice Sera. (Results are expressed as mean + SD (n= 3). Values with **, * are significant at p
    Figure Legend Snippet: Effect of HBsAg-Chitin formulation on IgG titre of Mice Sera. (Results are expressed as mean + SD (n= 3). Values with **, * are significant at p

    Techniques Used: Mouse Assay

    Effect HBsAg-Chitin formulation on IgG2a titre in Mice Sera. (Results are expressed as mean + SD (n= 3). Values with **, * are significant atp
    Figure Legend Snippet: Effect HBsAg-Chitin formulation on IgG2a titre in Mice Sera. (Results are expressed as mean + SD (n= 3). Values with **, * are significant atp

    Techniques Used: Mouse Assay

    Effect of HBsAg-Chitin formulation on IgG1 titre of Mice Sera. (Results are expressed as mean + SD (n= 3). Values with **, * are significant atp
    Figure Legend Snippet: Effect of HBsAg-Chitin formulation on IgG1 titre of Mice Sera. (Results are expressed as mean + SD (n= 3). Values with **, * are significant atp

    Techniques Used: Mouse Assay

    14) Product Images from "One-step generation of monoclonal B cell receptor mice capable of isotype switching and somatic hypermutation"

    Article Title: One-step generation of monoclonal B cell receptor mice capable of isotype switching and somatic hypermutation

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20172064

    Isotype switching and plasmablast differentiation in Igh CGG/+ mice in vitro. (a) In vitro isotype switching. Purified Igh CGG/+ (CD45.2/2) and WT (CD45.1/1) B cells were cocultured in vitro with LPS and IL-4 to induce isotype switching to IgG 1 . Histogram shows IgG 1 staining by flow cytometry on day 3 of culture. Data are from three independent experiments and are quantified in the graph; proportions of IgG 1 + cells in Igh CGG/+ and WT B cells from the same experiment are connected by a line. P = 0.74 (unpaired t test). (b) AFC cell formation (as defined by CD138 expression) and IgG 1 secretion in purified B cells stimulated with LPS and IL-4 in vitro. Cells were presorted as in Fig. 3 a , and non–CGG-binding B cells were excluded from Igh CGG/+ cultures. Histograms are representative of, and graphs show pooled data for two independent experiments. n = 3 mice for WT and n = 6 mice for Igh CGG/+ CGG binding B cells. P = 0.33 for CD138 expression and P = 0.37 for supernatant IgG 1 . Numbers within flow plots indicate percentage of cells in the designated gate.
    Figure Legend Snippet: Isotype switching and plasmablast differentiation in Igh CGG/+ mice in vitro. (a) In vitro isotype switching. Purified Igh CGG/+ (CD45.2/2) and WT (CD45.1/1) B cells were cocultured in vitro with LPS and IL-4 to induce isotype switching to IgG 1 . Histogram shows IgG 1 staining by flow cytometry on day 3 of culture. Data are from three independent experiments and are quantified in the graph; proportions of IgG 1 + cells in Igh CGG/+ and WT B cells from the same experiment are connected by a line. P = 0.74 (unpaired t test). (b) AFC cell formation (as defined by CD138 expression) and IgG 1 secretion in purified B cells stimulated with LPS and IL-4 in vitro. Cells were presorted as in Fig. 3 a , and non–CGG-binding B cells were excluded from Igh CGG/+ cultures. Histograms are representative of, and graphs show pooled data for two independent experiments. n = 3 mice for WT and n = 6 mice for Igh CGG/+ CGG binding B cells. P = 0.33 for CD138 expression and P = 0.37 for supernatant IgG 1 . Numbers within flow plots indicate percentage of cells in the designated gate.

    Techniques Used: Mouse Assay, In Vitro, Purification, Staining, Flow Cytometry, Cytometry, Expressing, Binding Assay

    CSR, SHM, and antigen-driven selection in Igh CGG/+ B cells. (a) Igh CGG/+ B cells (CD45.2/2) were adoptively transferred into WT (CD45.1/1) recipients, which were immunized 1 d later with CGG in alum. CSR to IgG 1 was determined in GC B cells (CD19 + TCRβ – Fas + CD38 lo ) from donor (CD45.2/2) and recipient (CD45.1/1) mice by flow cytometry. Graph shows percentage of IgG 1 + GC B cells in five mice from two independent experiments. Proportions of switched cells in the same mouse are connected by a line. P = 0.23 (unpaired t test). Numbers within flow plots indicate percentage of cells in the designated gate. (b) Left: Serum titer of IgG 1 + /human Cκ + Ig in three mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG. Graph shows data for three mice from two independent experiments. Right: Western blot for human Igκ in serum of mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG 14 d after immunization. NR, nonreducing conditions; R, reducing conditions (DTT). Blot is representative of two experiments. (c–f) GC B cells from adoptive transfer experiments as in panel a were single sorted, and the entire Igh CGG/+ allele was PCR-amplified and sequenced. (c) Nucleotide mutation frequencies along the Igh CGG/+ locus, calculated as the fraction of times a particular nucleotide was mutated from the original sequence. (d) Total mutation frequency per region (normalized to sequence length). Each symbol represents one cell. (e) Number of nucleotide mutations in VDJ H and VJκ in Igh CGG/+ GC B cells compared with WT GC B cells carrying the same rearrangement selected by CGG immunization, as reported previously ( Tas et al., 2016 ; only data from LN#2 GC2 in Fig. 4 from Tas et al., 2016 are analyzed). Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. **, P ≤ 0.001; ****, P ≤ 0.0001 (unpaired t test). (f) Comparison of nucleotide mutation patterns between Igh CGG/+ B cells and WT GC B cells carrying the same rearrangement. Mutation patterns are shown for FR1 to FR4 regions of IgH and CDR1 to FR4 regions of Igκ. Shared nucleotide mutation positions are shown in red. Asterisk indicates a C119 > G mutation, which confers an approximately eightfold gain in affinity. Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. (g) Reconstructed phylogeny of Igh CGG/+ GC B cell VJ Igκ sequences from multiple GCs in one LN. Clades of expanded clones are indicated in red, green, and blue. Cells containing the high-affinity C119 > G mutation are indicated in green.
    Figure Legend Snippet: CSR, SHM, and antigen-driven selection in Igh CGG/+ B cells. (a) Igh CGG/+ B cells (CD45.2/2) were adoptively transferred into WT (CD45.1/1) recipients, which were immunized 1 d later with CGG in alum. CSR to IgG 1 was determined in GC B cells (CD19 + TCRβ – Fas + CD38 lo ) from donor (CD45.2/2) and recipient (CD45.1/1) mice by flow cytometry. Graph shows percentage of IgG 1 + GC B cells in five mice from two independent experiments. Proportions of switched cells in the same mouse are connected by a line. P = 0.23 (unpaired t test). Numbers within flow plots indicate percentage of cells in the designated gate. (b) Left: Serum titer of IgG 1 + /human Cκ + Ig in three mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG. Graph shows data for three mice from two independent experiments. Right: Western blot for human Igκ in serum of mice adoptively transferred with Igh CGG/+ B cells and immunized with CGG 14 d after immunization. NR, nonreducing conditions; R, reducing conditions (DTT). Blot is representative of two experiments. (c–f) GC B cells from adoptive transfer experiments as in panel a were single sorted, and the entire Igh CGG/+ allele was PCR-amplified and sequenced. (c) Nucleotide mutation frequencies along the Igh CGG/+ locus, calculated as the fraction of times a particular nucleotide was mutated from the original sequence. (d) Total mutation frequency per region (normalized to sequence length). Each symbol represents one cell. (e) Number of nucleotide mutations in VDJ H and VJκ in Igh CGG/+ GC B cells compared with WT GC B cells carrying the same rearrangement selected by CGG immunization, as reported previously ( Tas et al., 2016 ; only data from LN#2 GC2 in Fig. 4 from Tas et al., 2016 are analyzed). Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. **, P ≤ 0.001; ****, P ≤ 0.0001 (unpaired t test). (f) Comparison of nucleotide mutation patterns between Igh CGG/+ B cells and WT GC B cells carrying the same rearrangement. Mutation patterns are shown for FR1 to FR4 regions of IgH and CDR1 to FR4 regions of Igκ. Shared nucleotide mutation positions are shown in red. Asterisk indicates a C119 > G mutation, which confers an approximately eightfold gain in affinity. Data from two transfers of Igh CGG/+ B cells are pooled and compared with the WT. (g) Reconstructed phylogeny of Igh CGG/+ GC B cell VJ Igκ sequences from multiple GCs in one LN. Clades of expanded clones are indicated in red, green, and blue. Cells containing the high-affinity C119 > G mutation are indicated in green.

    Techniques Used: Selection, Mouse Assay, Flow Cytometry, Cytometry, Western Blot, Adoptive Transfer Assay, Polymerase Chain Reaction, Amplification, Mutagenesis, Sequencing, Clone Assay

    15) Product Images from "Identification of T helper (Th)1- and Th2-associated antigens of Cryptococcus neoformans in a murine model of pulmonary infection"

    Article Title: Identification of T helper (Th)1- and Th2-associated antigens of Cryptococcus neoformans in a murine model of pulmonary infection

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-21039-z

    IgG2a-immunoreactive proteins from Cryptococcus neoformans were detected with sera from representative infected and naïve wild-type, IL-4Rα-deficient, and IL-12-deficient mice. Whole cell proteins of C. neoformans strain 1841 separated by 2D electrophoresis were transferred to nitrocellulose membranes, which were thereafter incubated with sera from infected and naïve wild-type and IL-4Rα-deficient mice diluted 1:1,000. IgG2a-immunoreactive proteins were detected using sera from an infected ( a ) and naïve ( b ) wild-type mouse, an infected ( c ) and naïve ( d ) IL-4Rα-deficient mouse, and an infected ( e ) and naïve ( f ) IL-12-deficient mouse. Protein abundance as shown in the Coomassie staining did not correlate with the strength of the immunoreactive signal (Fig. 4 ). Only the spots that could be mapped on Coomassie-stained gels were numbered in the blot images. Bold, underlined numbers mark IgG2a-reactive, C. neoformans -specific spots, while light, underlined numbers indicate IgG2a-reactive but C. neoformans -unspecific spots. Italic numbers mark spots reactive with both IgG1 and IgG2a antibodies. Images were cropped to improve clarity. Full-length blots without numbered protein spots are presented in Supplementary Fig. 3 . Abbreviation: MM = molecular mass.
    Figure Legend Snippet: IgG2a-immunoreactive proteins from Cryptococcus neoformans were detected with sera from representative infected and naïve wild-type, IL-4Rα-deficient, and IL-12-deficient mice. Whole cell proteins of C. neoformans strain 1841 separated by 2D electrophoresis were transferred to nitrocellulose membranes, which were thereafter incubated with sera from infected and naïve wild-type and IL-4Rα-deficient mice diluted 1:1,000. IgG2a-immunoreactive proteins were detected using sera from an infected ( a ) and naïve ( b ) wild-type mouse, an infected ( c ) and naïve ( d ) IL-4Rα-deficient mouse, and an infected ( e ) and naïve ( f ) IL-12-deficient mouse. Protein abundance as shown in the Coomassie staining did not correlate with the strength of the immunoreactive signal (Fig. 4 ). Only the spots that could be mapped on Coomassie-stained gels were numbered in the blot images. Bold, underlined numbers mark IgG2a-reactive, C. neoformans -specific spots, while light, underlined numbers indicate IgG2a-reactive but C. neoformans -unspecific spots. Italic numbers mark spots reactive with both IgG1 and IgG2a antibodies. Images were cropped to improve clarity. Full-length blots without numbered protein spots are presented in Supplementary Fig. 3 . Abbreviation: MM = molecular mass.

    Techniques Used: Infection, Mouse Assay, Two-Dimensional Gel Electrophoresis, Incubation, Staining

    Frequency of cryptococcal proteins, reactive with IgG1, IgG2a, or with IgG1 and IgG2a antibodies in Cryptococcus neoformans -infected mice of different genotypes. Immunoreactive cryptococcal proteins were identified using sera from infected mice of different genotypes. Proteins were grouped according to their reactivity with IgG1, IgG2a, or with IgG1 and IgG2a antibodies. The isotype which showed reactivity in the individual animal is indicated by plain bars (IgG1) or hatched bars (IgG2a). Three IgG2a-reactive proteins reacted exclusively with sera from infected mice, while one protein also showed reactivity with sera from naïve mice (marked with an asterisk (*), see also Fig. 3 ). Sera from IL-4Rα-deficient mice were not tested by 2D analysis for IgG1-reactive proteins, as no immunoreactive protein bands for this isotype could be detected when investigating these sera in 1D analysis (Supplementary Fig. S1 ). Sera from infected animals were taken from at least three independent experiments in late infection state (at least 56 days post infection). Abbreviations: Glyceraldehyde-3-phosphate dehyd. = Glyceraldehyde-3-phosphate dehydrogenase; Thioredoxin-dependent peroxide reduct. = Thioredoxin-dependent peroxide reductase.
    Figure Legend Snippet: Frequency of cryptococcal proteins, reactive with IgG1, IgG2a, or with IgG1 and IgG2a antibodies in Cryptococcus neoformans -infected mice of different genotypes. Immunoreactive cryptococcal proteins were identified using sera from infected mice of different genotypes. Proteins were grouped according to their reactivity with IgG1, IgG2a, or with IgG1 and IgG2a antibodies. The isotype which showed reactivity in the individual animal is indicated by plain bars (IgG1) or hatched bars (IgG2a). Three IgG2a-reactive proteins reacted exclusively with sera from infected mice, while one protein also showed reactivity with sera from naïve mice (marked with an asterisk (*), see also Fig. 3 ). Sera from IL-4Rα-deficient mice were not tested by 2D analysis for IgG1-reactive proteins, as no immunoreactive protein bands for this isotype could be detected when investigating these sera in 1D analysis (Supplementary Fig. S1 ). Sera from infected animals were taken from at least three independent experiments in late infection state (at least 56 days post infection). Abbreviations: Glyceraldehyde-3-phosphate dehyd. = Glyceraldehyde-3-phosphate dehydrogenase; Thioredoxin-dependent peroxide reduct. = Thioredoxin-dependent peroxide reductase.

    Techniques Used: Infection, Mouse Assay

    Total and C. neoformans -specific levels of Th2-dependent IgG1 and IgE predominate in sera from susceptible wild-type (WT) and IL-12-deficient mice with pulmonary cryptococcosis. Total IgG1 ( a ) and IgE ( b ) levels increased significantly after infection with C. neoformans for most genotypes, whereas IgG2a levels ( c ) were not influenced by C. neoformans . Compared to wild-type mice, IL-4Rα-deficient mice had markedly lower levels of immunoglobulin (Ig)G1 ( a ), no production of IgE ( b ) and similar levels of total IgG2a ( c ). Opposite, IL-12-deficient mice showed similar IgG1 and significantly higher IgE levels ( a , b ), while reduced levels of IgG2a ( c ), compared to wild-type mice. Titers of C. neoformans -specific IgG2a ( d ) and IgG1 ( e ) antibodies in infected (infec.) mice reflected the distribution observed for total immunoglobulin levels. C. neoformans -specific IgG1 and IgG2a antibodies were absent in sera of naïve mice. A strong correlation was observed between total and C. neoformans -specific IgG1 levels ( f ) but not between total and C. neoformans -specific IgG2a levels ( g ) in infected mice of all genotypes. Each spot represents a serum sample of an individual mouse (2 to 14 animals per group from at least two independent experiments) with the line indicating the median. Serum samples were obtained from mice intranasally infected with 500 colony forming units of C. neoformans strain 1841 after about 56 days post infection. Statistical significance determined by Mann-Whitney U test is shown as following: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. Correlation was determined by nonparametric Spearman’s correlation test.
    Figure Legend Snippet: Total and C. neoformans -specific levels of Th2-dependent IgG1 and IgE predominate in sera from susceptible wild-type (WT) and IL-12-deficient mice with pulmonary cryptococcosis. Total IgG1 ( a ) and IgE ( b ) levels increased significantly after infection with C. neoformans for most genotypes, whereas IgG2a levels ( c ) were not influenced by C. neoformans . Compared to wild-type mice, IL-4Rα-deficient mice had markedly lower levels of immunoglobulin (Ig)G1 ( a ), no production of IgE ( b ) and similar levels of total IgG2a ( c ). Opposite, IL-12-deficient mice showed similar IgG1 and significantly higher IgE levels ( a , b ), while reduced levels of IgG2a ( c ), compared to wild-type mice. Titers of C. neoformans -specific IgG2a ( d ) and IgG1 ( e ) antibodies in infected (infec.) mice reflected the distribution observed for total immunoglobulin levels. C. neoformans -specific IgG1 and IgG2a antibodies were absent in sera of naïve mice. A strong correlation was observed between total and C. neoformans -specific IgG1 levels ( f ) but not between total and C. neoformans -specific IgG2a levels ( g ) in infected mice of all genotypes. Each spot represents a serum sample of an individual mouse (2 to 14 animals per group from at least two independent experiments) with the line indicating the median. Serum samples were obtained from mice intranasally infected with 500 colony forming units of C. neoformans strain 1841 after about 56 days post infection. Statistical significance determined by Mann-Whitney U test is shown as following: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. Correlation was determined by nonparametric Spearman’s correlation test.

    Techniques Used: Mouse Assay, Infection, MANN-WHITNEY

    Protein profile of Cryptococcus neoformans with indicated immunoreactive protein spots. Whole cell proteins of C. neoformans strain 1841 were separated by isoelectric point and molecular weight. After 2D gel electrophoresis, gels were stained with Coomassie Brilliant Blue G250. Numbered spots in the stained gel represent all antigenic proteins that were identified in this study. Bold non-underlined and bold underlined numbers indicate IgG1- and IgG2a-immunoreactive proteins, respectively. The spots in italic were reactive with both isotypes as shown in Figures 2 and 3 . Light underlined numbers indicate IgG2a-immunoreactive proteins, which were not specific for C. neoformans . Abbreviation: MM = molecular mass.
    Figure Legend Snippet: Protein profile of Cryptococcus neoformans with indicated immunoreactive protein spots. Whole cell proteins of C. neoformans strain 1841 were separated by isoelectric point and molecular weight. After 2D gel electrophoresis, gels were stained with Coomassie Brilliant Blue G250. Numbered spots in the stained gel represent all antigenic proteins that were identified in this study. Bold non-underlined and bold underlined numbers indicate IgG1- and IgG2a-immunoreactive proteins, respectively. The spots in italic were reactive with both isotypes as shown in Figures 2 and 3 . Light underlined numbers indicate IgG2a-immunoreactive proteins, which were not specific for C. neoformans . Abbreviation: MM = molecular mass.

    Techniques Used: Molecular Weight, Two-Dimensional Gel Electrophoresis, Electrophoresis, Staining

    IgG1-immunoreactive proteins from Cryptococcus neoformans were detected with sera from representative infected but not naïve wild-type and IL-12-deficient mice. Whole cell proteins of C. neoformans strain 1841 separated by 2D electrophoresis were transferred to nitrocellulose membranes, which were thereafter incubated with sera from infected and naïve wild-type and gene-deficient mice diluted 1:1,000. IgG1-immunoreactive proteins were detected using sera from an infected wild-type ( a ), a naïve wild-type ( b ), an infected IL-12-deficient ( c ), and a naïve IL-12-deficient ( d ) mouse. Protein abundance as shown in the Coomassie staining did not correlate with the strength of the immunoreactive signal (Fig. 4 ). Only the spots that could be mapped on Coomassie-stained gels were numbered in the blot images. Bold numbers indicate strictly IgG1-reactive proteins while italic numbers mark proteins reactive with both, IgG1 and IgG2a antibodies. Images were cropped to improve clarity. Full-length blots without numbered protein spots are presented in Supplementary Fig. 2 . Abbreviation: MM = molecular mass.
    Figure Legend Snippet: IgG1-immunoreactive proteins from Cryptococcus neoformans were detected with sera from representative infected but not naïve wild-type and IL-12-deficient mice. Whole cell proteins of C. neoformans strain 1841 separated by 2D electrophoresis were transferred to nitrocellulose membranes, which were thereafter incubated with sera from infected and naïve wild-type and gene-deficient mice diluted 1:1,000. IgG1-immunoreactive proteins were detected using sera from an infected wild-type ( a ), a naïve wild-type ( b ), an infected IL-12-deficient ( c ), and a naïve IL-12-deficient ( d ) mouse. Protein abundance as shown in the Coomassie staining did not correlate with the strength of the immunoreactive signal (Fig. 4 ). Only the spots that could be mapped on Coomassie-stained gels were numbered in the blot images. Bold numbers indicate strictly IgG1-reactive proteins while italic numbers mark proteins reactive with both, IgG1 and IgG2a antibodies. Images were cropped to improve clarity. Full-length blots without numbered protein spots are presented in Supplementary Fig. 2 . Abbreviation: MM = molecular mass.

    Techniques Used: Infection, Mouse Assay, Two-Dimensional Gel Electrophoresis, Incubation, Staining

    16) Product Images from "IL-23, not IL-12, Directs Autoimmunity to the Goodpasture Antigen"

    Article Title: IL-23, not IL-12, Directs Autoimmunity to the Goodpasture Antigen

    Journal: Journal of the American Society of Nephrology : JASN

    doi: 10.1681/ASN.2008080891

    Humoral responses and anti-α3(IV)NC1 autoantibody production. (A and B) α3(IV)NC1-specific CD19 + proliferation (A) and activation (B) were decreased in IL-12p40 −/− and in IL-23p19 −/− mice compared with WT or IL-12p35 −/− mice. A and B show both representative FACS plots from single animals and a graph that summarizes data from all animals. Numbers within the FACS plot quadrants represent percentage of all cells analyzed. Numbers in top right quadrants represent the percentage of CD19 + cells that were either BrdU + or CD69 + . (C) Early α3(IV)NC1-specific IgG autoantibody titers were reduced in IL-12p40 −/− and in IL-23p19 −/− mice when compared with WT or IL-12p35 −/− mice (sera dilution 1:2000). (D and E) The reduction in antibody titers persisted during established disease (D) and included α3(IV)NC1-specific IgG1 (1:10,000), IgG2b (1:10,000), and IgG3 (1:250) titers (E). (D) IgG and IgG2b titers were modestly reduced in IL-12p35 −/− mice during established disease. * P
    Figure Legend Snippet: Humoral responses and anti-α3(IV)NC1 autoantibody production. (A and B) α3(IV)NC1-specific CD19 + proliferation (A) and activation (B) were decreased in IL-12p40 −/− and in IL-23p19 −/− mice compared with WT or IL-12p35 −/− mice. A and B show both representative FACS plots from single animals and a graph that summarizes data from all animals. Numbers within the FACS plot quadrants represent percentage of all cells analyzed. Numbers in top right quadrants represent the percentage of CD19 + cells that were either BrdU + or CD69 + . (C) Early α3(IV)NC1-specific IgG autoantibody titers were reduced in IL-12p40 −/− and in IL-23p19 −/− mice when compared with WT or IL-12p35 −/− mice (sera dilution 1:2000). (D and E) The reduction in antibody titers persisted during established disease (D) and included α3(IV)NC1-specific IgG1 (1:10,000), IgG2b (1:10,000), and IgG3 (1:250) titers (E). (D) IgG and IgG2b titers were modestly reduced in IL-12p35 −/− mice during established disease. * P

    Techniques Used: Activation Assay, Mouse Assay, FACS

    17) Product Images from "Dendritic cell-bound IgE functions to restrain allergic inflammation at mucosal sites"

    Article Title: Dendritic cell-bound IgE functions to restrain allergic inflammation at mucosal sites

    Journal: Mucosal immunology

    doi: 10.1038/mi.2014.85

    Intestinal DCs express FcεRI and allergic sensitization increases the DC-bound IgE pool of IgE R -TG animals. ( a) FcεRI expression (in red) in the small intestine of IgE R -TG mice is found on CD11c + DCs (in blue). CD11c + DCs from IgE R -TG are GFP positive because the transgenic construct contains an IRES-GPF reporter element. (b) FcεRI expression on CD11c + DCs in the human small intestine. Human FcεRIα (green, first panel), CD11c (red, second panel). Mast cells as depicted by c-kit are not found in non-inflamed human tissue using immunofluorescence (row two, red). DAPI-stain for nuclei (blue). (c) and (d) Analysis of OVA-specific IgE and IgG1 levels in serum of sensitized mice before antigen challenge. (e) and (f) Sensitization and antigen challenge increases the amount of surface-bound IgE on DCs of IgE R -TG mice but not WT animals. DCs were isolated from the spleens and mesenteric lymph nodes (MLNs) of OVA-challenged WT and IgE R -TG mice, respectively. Sensitized mice are indicated by (+), non-sensitized mice by (−). Symbols are representative of individual mice (n=2).
    Figure Legend Snippet: Intestinal DCs express FcεRI and allergic sensitization increases the DC-bound IgE pool of IgE R -TG animals. ( a) FcεRI expression (in red) in the small intestine of IgE R -TG mice is found on CD11c + DCs (in blue). CD11c + DCs from IgE R -TG are GFP positive because the transgenic construct contains an IRES-GPF reporter element. (b) FcεRI expression on CD11c + DCs in the human small intestine. Human FcεRIα (green, first panel), CD11c (red, second panel). Mast cells as depicted by c-kit are not found in non-inflamed human tissue using immunofluorescence (row two, red). DAPI-stain for nuclei (blue). (c) and (d) Analysis of OVA-specific IgE and IgG1 levels in serum of sensitized mice before antigen challenge. (e) and (f) Sensitization and antigen challenge increases the amount of surface-bound IgE on DCs of IgE R -TG mice but not WT animals. DCs were isolated from the spleens and mesenteric lymph nodes (MLNs) of OVA-challenged WT and IgE R -TG mice, respectively. Sensitized mice are indicated by (+), non-sensitized mice by (−). Symbols are representative of individual mice (n=2).

    Techniques Used: Expressing, Mouse Assay, Transgenic Assay, Construct, Immunofluorescence, Staining, Isolation

    18) Product Images from "LatY136F knock-in mouse model for human IgG4-related disease"

    Article Title: LatY136F knock-in mouse model for human IgG4-related disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0198417

    Immunostainings for plasma cells, T cells and macrophages in tissue lesions of 6-week-old Lat Y136F knock-in mice. Numerous CD138-positive plasma cells (A; kidney, magnification: 200×), CD4-positive cells (B; kidney, magnification: 400×), and CD8-positive cells (C; kidney, D; magnification: 400×) were detected. F4/80-positive cells also infiltrated the kidney (D; magnification: 400×). IgG- and IgG1-positive cells infiltrated the kidneys (E and F; magnification: 400×). Immunostaining using consecutive sections showed mostly CD138 positive cells (G; kidney, magnification: 400×) were IgG1-positive (H; kidney, magnification: 400×). Bars in each panel showed 40 μm. Arrows in B-F indicated representative cells.
    Figure Legend Snippet: Immunostainings for plasma cells, T cells and macrophages in tissue lesions of 6-week-old Lat Y136F knock-in mice. Numerous CD138-positive plasma cells (A; kidney, magnification: 200×), CD4-positive cells (B; kidney, magnification: 400×), and CD8-positive cells (C; kidney, D; magnification: 400×) were detected. F4/80-positive cells also infiltrated the kidney (D; magnification: 400×). IgG- and IgG1-positive cells infiltrated the kidneys (E and F; magnification: 400×). Immunostaining using consecutive sections showed mostly CD138 positive cells (G; kidney, magnification: 400×) were IgG1-positive (H; kidney, magnification: 400×). Bars in each panel showed 40 μm. Arrows in B-F indicated representative cells.

    Techniques Used: Knock-In, Mouse Assay, Immunostaining

    19) Product Images from "Helicobacter bilis triggers persistent immune reactivity to antigens derived from the commensal bacteria in gnotobiotic C3H/HeN mice"

    Article Title: Helicobacter bilis triggers persistent immune reactivity to antigens derived from the commensal bacteria in gnotobiotic C3H/HeN mice

    Journal:

    doi: 10.1136/gut.2006.099242

    Figure 4 Measurement of serum IgG1 and IgG2a antibody responses to H bilis antigen collected at various time points following colonisation of defined flora mice with H bilis as described in fig 1. The antibody response was determined
    Figure Legend Snippet: Figure 4 Measurement of serum IgG1 and IgG2a antibody responses to H bilis antigen collected at various time points following colonisation of defined flora mice with H bilis as described in fig 1. The antibody response was determined

    Techniques Used: Mouse Assay

    Figure 3 Measurement of serum immunoglobin (Ig) G1 and IgG2a responses to altered Schaedler flora antigens for samples collected from non‐infected (NI) and H bilis ‐colonised defined flora mice as described in fig 1. The
    Figure Legend Snippet: Figure 3 Measurement of serum immunoglobin (Ig) G1 and IgG2a responses to altered Schaedler flora antigens for samples collected from non‐infected (NI) and H bilis ‐colonised defined flora mice as described in fig 1. The

    Techniques Used: Infection, Mouse Assay

    20) Product Images from "Cathepsin B-Deficient Mice Resolve Leishmania major Inflammation Faster in a T Cell-Dependent Manner"

    Article Title: Cathepsin B-Deficient Mice Resolve Leishmania major Inflammation Faster in a T Cell-Dependent Manner

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0004716

    Protective local cytokine responses develop faster and resolve sooner in CatB -/- mice. WT and CatB -/- were subcutaneously inoculated with 3x10 6 stationary phase promastigotes of L . major in the footpads and experimental read-outs were measured weekly as indicated. Expression levels of IFNγ (A) , IL-4 (B) , IL-10 (C) and iNOS (D) transcripts were determined in lymph nodes of L . major -infected WT (filled bars) and CatB -/- (empty bars) mice by RT-PCR. Values are normalized to the expression of the HPRT gene. Levels of SLA-specific IgG1 (E) and IgG2a (F) were determined by ELISA on serum samples harvested from L . major -infected WT (filled bars) and CatB -/- (empty bars) mice. Data depicted represent the mean and SEM of at least 3 independent experiments with n ≥ 4 mice/group/time point. * p
    Figure Legend Snippet: Protective local cytokine responses develop faster and resolve sooner in CatB -/- mice. WT and CatB -/- were subcutaneously inoculated with 3x10 6 stationary phase promastigotes of L . major in the footpads and experimental read-outs were measured weekly as indicated. Expression levels of IFNγ (A) , IL-4 (B) , IL-10 (C) and iNOS (D) transcripts were determined in lymph nodes of L . major -infected WT (filled bars) and CatB -/- (empty bars) mice by RT-PCR. Values are normalized to the expression of the HPRT gene. Levels of SLA-specific IgG1 (E) and IgG2a (F) were determined by ELISA on serum samples harvested from L . major -infected WT (filled bars) and CatB -/- (empty bars) mice. Data depicted represent the mean and SEM of at least 3 independent experiments with n ≥ 4 mice/group/time point. * p

    Techniques Used: Mouse Assay, Expressing, Infection, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

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    SouthernBiotech alkaline phosphatase labeled goat anti mouse igg1
    mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C <t>IgG1</t> control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.
    Alkaline Phosphatase Labeled Goat Anti Mouse Igg1, supplied by SouthernBiotech, 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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    SouthernBiotech mouse igg isotype control
    A glycopeptide epitope on lung cancer cell surface is preferably recognized by mAb 16A. Lung adenocarcinoma cell lines, NCI-H1395, HCC4019, H838, H1573, H1703, H2030, and H3255 were studied by flow cytometry staining. Monoclonal antibodies 14A, which binds to MUC1 peptide part only (RPAPGSTAPPAHG); 16A, which binds to MUC1 glycopeptide RPAPGS(GalNAc)TAPPAHG; and B72.3, which binds to sugars only (clustered Tn antigen), were used as primary antibodies. Goat anti-mouse <t>IgG</t> (Allophycocyanin-conjugated), and mouse IgG isotype control were from Southern Biotech (Birmingham, AL, USA).
    Mouse Igg Isotype Control, supplied by SouthernBiotech, 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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    SouthernBiotech anti igg2b
    Impaired B cell responses in TC10 KO animals. ( A ) Serum was collected from naive WT and TC10 KO animals, and baseline titers of IgM, <t>IgG2b,</t> IgG2c, IgG3, and IgA were measured by sandwich ELISA. ( B – E ) WT and TC10 KO animals were infected with 10 4 PFU VACV (B and C) or 2.10 2 Influenza virus (D and E) by intra footpad (B and C) or intranasal injection (D and E), and the draining popliteal (B and C) or mediastinal (D and E) LNs were isolated 7 (B and C) or 9 (D and E) d later. Germinal center B cells (CD95 + GL-7 + (B and D) and plasma cells [CD138 + IgD lo (C and E)] were analyzed by flow cytometry. Quantifications are shown on the panels on the right and show the percentage of B cells in the indicated gates. ( F and G ) WT and TC10 KO mice were immunized with NP-KLH precipitated in Alum, and serum samples collected weekly for 28 d. NP-specific IgM and IgG3 (F) titers were measured, as well as total IgG titers (G). ( H ) ELISA analysis showing affinity maturation (expressed as the ratio of NP3 to NP23 titers) of total IgG in WT and TC10 KO animals. Data are from one representative of two to three independent experiments with at least three animals in each group. Student t test (ns: p > 0.05, * p
    Anti Igg2b, supplied by SouthernBiotech, 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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    SouthernBiotech goat anti mouse collagen type i
    Distribution of <t>HMGB1</t> is in accordance with that of <t>type</t> I collagen in fibrotic mice. Expression levels of HMGB1 were detected using immunofluorescence. Representative HMGB1 staining of all groups is shown (Original magnification, ×200). HMGB1, high-mobility group box 1; Coll-1, type I collagen.
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    Image Search Results


    mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C IgG1 control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.

    Journal: The Journal of Biological Chemistry

    Article Title: A Monoclonal Antibody to Cryptococcus neoformans Glucuronoxylomannan Manifests Hydrolytic Activity for Both Peptides and Polysaccharides *

    doi: 10.1074/jbc.M116.767582

    Figure Lengend Snippet: mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C IgG1 control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.

    Article Snippet: Completion of the assay was performed by successively adding the anti-GXM IgG1 mAb 18B7 at 10 μg/ml and then alkaline phosphatase-labeled goat anti-mouse IgG1 at 1 μg/ml (SouthernBiotech).

    Techniques: Flow Cytometry, Cytometry, Construct, Incubation

    A glycopeptide epitope on lung cancer cell surface is preferably recognized by mAb 16A. Lung adenocarcinoma cell lines, NCI-H1395, HCC4019, H838, H1573, H1703, H2030, and H3255 were studied by flow cytometry staining. Monoclonal antibodies 14A, which binds to MUC1 peptide part only (RPAPGSTAPPAHG); 16A, which binds to MUC1 glycopeptide RPAPGS(GalNAc)TAPPAHG; and B72.3, which binds to sugars only (clustered Tn antigen), were used as primary antibodies. Goat anti-mouse IgG (Allophycocyanin-conjugated), and mouse IgG isotype control were from Southern Biotech (Birmingham, AL, USA).

    Journal: International Journal of Oncology

    Article Title: Molecular basis of antibody binding to mucin glycopeptides in lung cancer

    doi: 10.3892/ijo.2015.3302

    Figure Lengend Snippet: A glycopeptide epitope on lung cancer cell surface is preferably recognized by mAb 16A. Lung adenocarcinoma cell lines, NCI-H1395, HCC4019, H838, H1573, H1703, H2030, and H3255 were studied by flow cytometry staining. Monoclonal antibodies 14A, which binds to MUC1 peptide part only (RPAPGSTAPPAHG); 16A, which binds to MUC1 glycopeptide RPAPGS(GalNAc)TAPPAHG; and B72.3, which binds to sugars only (clustered Tn antigen), were used as primary antibodies. Goat anti-mouse IgG (Allophycocyanin-conjugated), and mouse IgG isotype control were from Southern Biotech (Birmingham, AL, USA).

    Article Snippet: Goat anti-mouse IgG (Allophycocyanin-conjugated), and mouse IgG isotype control were from Southern Biotech (Birmingham, AL, USA).

    Techniques: Flow Cytometry, Cytometry, Staining

    Impaired B cell responses in TC10 KO animals. ( A ) Serum was collected from naive WT and TC10 KO animals, and baseline titers of IgM, IgG2b, IgG2c, IgG3, and IgA were measured by sandwich ELISA. ( B – E ) WT and TC10 KO animals were infected with 10 4 PFU VACV (B and C) or 2.10 2 Influenza virus (D and E) by intra footpad (B and C) or intranasal injection (D and E), and the draining popliteal (B and C) or mediastinal (D and E) LNs were isolated 7 (B and C) or 9 (D and E) d later. Germinal center B cells (CD95 + GL-7 + (B and D) and plasma cells [CD138 + IgD lo (C and E)] were analyzed by flow cytometry. Quantifications are shown on the panels on the right and show the percentage of B cells in the indicated gates. ( F and G ) WT and TC10 KO mice were immunized with NP-KLH precipitated in Alum, and serum samples collected weekly for 28 d. NP-specific IgM and IgG3 (F) titers were measured, as well as total IgG titers (G). ( H ) ELISA analysis showing affinity maturation (expressed as the ratio of NP3 to NP23 titers) of total IgG in WT and TC10 KO animals. Data are from one representative of two to three independent experiments with at least three animals in each group. Student t test (ns: p > 0.05, * p

    Journal: The Journal of Immunology Author Choice

    Article Title: The Small Rho GTPase TC10 Modulates B Cell Immune Responses

    doi: 10.4049/jimmunol.1602167

    Figure Lengend Snippet: Impaired B cell responses in TC10 KO animals. ( A ) Serum was collected from naive WT and TC10 KO animals, and baseline titers of IgM, IgG2b, IgG2c, IgG3, and IgA were measured by sandwich ELISA. ( B – E ) WT and TC10 KO animals were infected with 10 4 PFU VACV (B and C) or 2.10 2 Influenza virus (D and E) by intra footpad (B and C) or intranasal injection (D and E), and the draining popliteal (B and C) or mediastinal (D and E) LNs were isolated 7 (B and C) or 9 (D and E) d later. Germinal center B cells (CD95 + GL-7 + (B and D) and plasma cells [CD138 + IgD lo (C and E)] were analyzed by flow cytometry. Quantifications are shown on the panels on the right and show the percentage of B cells in the indicated gates. ( F and G ) WT and TC10 KO mice were immunized with NP-KLH precipitated in Alum, and serum samples collected weekly for 28 d. NP-specific IgM and IgG3 (F) titers were measured, as well as total IgG titers (G). ( H ) ELISA analysis showing affinity maturation (expressed as the ratio of NP3 to NP23 titers) of total IgG in WT and TC10 KO animals. Data are from one representative of two to three independent experiments with at least three animals in each group. Student t test (ns: p > 0.05, * p

    Article Snippet: Biotinylated anti-IgM (553406; BD Biosciences), anti-IgG2b (Southern Biotech), IgG2c (Southern Biotech), or anti-IL6 (Clone MP5-32C11; BD Biosciences) Abs were used for detection.

    Techniques: Sandwich ELISA, Infection, Injection, Isolation, Flow Cytometry, Cytometry, Mouse Assay, Enzyme-linked Immunosorbent Assay

    Distribution of HMGB1 is in accordance with that of type I collagen in fibrotic mice. Expression levels of HMGB1 were detected using immunofluorescence. Representative HMGB1 staining of all groups is shown (Original magnification, ×200). HMGB1, high-mobility group box 1; Coll-1, type I collagen.

    Journal: Molecular Medicine Reports

    Article Title: Inhibition of the translocation and extracellular release of high-mobility group box 1 alleviates liver damage in fibrotic mice in response to D-galactosamine/lipopolysaccharide challenge

    doi: 10.3892/mmr.2016.5003

    Figure Lengend Snippet: Distribution of HMGB1 is in accordance with that of type I collagen in fibrotic mice. Expression levels of HMGB1 were detected using immunofluorescence. Representative HMGB1 staining of all groups is shown (Original magnification, ×200). HMGB1, high-mobility group box 1; Coll-1, type I collagen.

    Article Snippet: For immunofluorescence staining, the liver sections were fixed and stained with the following primary antibodies at 4°C overnight: Rabbit anti-mouse HMGB1 monoclonal antibody (1:100; cat. no ab79823, Abcam, Cambridge, MA, USA), goat anti-mouse collagen type I (1:200; cat. no. 1310-01, Southern Biotech, San Diego, CA, USA) and DAPI (EMD Millipore, Billerica, MA, USA).

    Techniques: Mouse Assay, Expressing, Immunofluorescence, Staining