biotinylated polyclonal goat f ab 2 anti igg2a  (SouthernBiotech)

 
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    Name:
    Goat Anti Rabbit IgG Fab Mouse Human SP ads UNLB
    Description:

    Catalog Number:
    4058-01
    Price:
    None
    Source:
    Pooled antisera from goats hyperimmunized with rabbit IgG
    Applications:
    Quality tested applications for relevant formats include -ELISA
    Format:
    UNLB (Unconjugated)
    Isotype:
    Goat IgG
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    Structured Review

    SouthernBiotech biotinylated polyclonal goat f ab 2 anti igg2a
    Goat Anti Rabbit IgG Fab Mouse Human SP ads UNLB

    https://www.bioz.com/result/biotinylated polyclonal goat f ab 2 anti igg2a/product/SouthernBiotech
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated polyclonal goat f ab 2 anti igg2a - by Bioz Stars, 2021-07
    80/100 stars

    Images

    1) Product Images from "Cross-reactive antigen expressed by B6 splenocytes drives receptor editing and MZ differentiation of IgG2a-reactive AM14 Vκ8 B cells"

    Article Title: Cross-reactive antigen expressed by B6 splenocytes drives receptor editing and MZ differentiation of IgG2a-reactive AM14 Vκ8 B cells

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

    doi: 10.4049/jimmunol.1900499

    B6 AM14 Vκ8 B cells have an anergic phenotype A. Serum IgG2a levels of AM14 Vκ8/Vκ8 BALB/c (○) and B6 (π) mice were measured by ELISA (n=5 BALB/c, n=11 B6, mean +/− SEM). B. Number of B220+ B cells in the spleen of AM14 Vκ8/Vκ8 BALB/c (white bar, n=19) and B6 (black bar, n=9). Mean +/− SEM is shown. C. Proliferation of B220-purified splenic B cells in response to 15 μg/ml goat anti-IgM F(ab’) 2 (left) or 1 μg/ml ODN1826 (right) was measured by 3 H-thymidine uptake after 30h (n=7 BALB/c and B6, mean +/− SEM). D. Distribution of CD23 hi CD21 lo FO and CD23 lo CD21 hi MZ B cells of B220 + AA4.1 − mature B cells in the spleen was ascertained by flow cytometry (representative flow plots are shown, summary plot of n=19 BALB/c, n=9 B6 is shown on the right, mean +/− SEM). E. CD23-purified splenic B cells from AM14 Vκ8/Vκ8 BALB/c (○) and B6 (π) mice were stimulated with anti-IgM (left) and ODN1826 (right) and proliferation was measured as in C (n=9 BALB/c, n=7 B6, mean +/− SEM). F. Surface expression levels of the AM14 Vκ8 BCR (4G7), IgM and IgD on AM14 Vκ8/Vκ8 BALB/c B cells (white bar) and AM14 Vκ8/Vκ8 B6 (black bar) were measured by flow cytometry. Mean fluorescent intensity (MFI) was calculated using FlowJo software. The mean +/− SEM of n=6 mice is shown. G. B cell subsets in the bone marrow of AM14 Vκ8/Vκ8 BALB/c and B6 mice were analyzed using flow cytometry according to Hardy fractions B-D (B220 low IgM neg ), E (B220 low IgM + ) and F (B220 hi IgM + ). A summary graph of the different subsets is shown on the right for AM14 Vκ8/Vκ8 BALB/c mice (white bar, n=16) and AM14 Vκ8/Vκ8 B6 mice (black bar, n=14). Mean +/− SEM is shown.
    Figure Legend Snippet: B6 AM14 Vκ8 B cells have an anergic phenotype A. Serum IgG2a levels of AM14 Vκ8/Vκ8 BALB/c (○) and B6 (π) mice were measured by ELISA (n=5 BALB/c, n=11 B6, mean +/− SEM). B. Number of B220+ B cells in the spleen of AM14 Vκ8/Vκ8 BALB/c (white bar, n=19) and B6 (black bar, n=9). Mean +/− SEM is shown. C. Proliferation of B220-purified splenic B cells in response to 15 μg/ml goat anti-IgM F(ab’) 2 (left) or 1 μg/ml ODN1826 (right) was measured by 3 H-thymidine uptake after 30h (n=7 BALB/c and B6, mean +/− SEM). D. Distribution of CD23 hi CD21 lo FO and CD23 lo CD21 hi MZ B cells of B220 + AA4.1 − mature B cells in the spleen was ascertained by flow cytometry (representative flow plots are shown, summary plot of n=19 BALB/c, n=9 B6 is shown on the right, mean +/− SEM). E. CD23-purified splenic B cells from AM14 Vκ8/Vκ8 BALB/c (○) and B6 (π) mice were stimulated with anti-IgM (left) and ODN1826 (right) and proliferation was measured as in C (n=9 BALB/c, n=7 B6, mean +/− SEM). F. Surface expression levels of the AM14 Vκ8 BCR (4G7), IgM and IgD on AM14 Vκ8/Vκ8 BALB/c B cells (white bar) and AM14 Vκ8/Vκ8 B6 (black bar) were measured by flow cytometry. Mean fluorescent intensity (MFI) was calculated using FlowJo software. The mean +/− SEM of n=6 mice is shown. G. B cell subsets in the bone marrow of AM14 Vκ8/Vκ8 BALB/c and B6 mice were analyzed using flow cytometry according to Hardy fractions B-D (B220 low IgM neg ), E (B220 low IgM + ) and F (B220 hi IgM + ). A summary graph of the different subsets is shown on the right for AM14 Vκ8/Vκ8 BALB/c mice (white bar, n=16) and AM14 Vκ8/Vκ8 B6 mice (black bar, n=14). Mean +/− SEM is shown.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Purification, Flow Cytometry, Expressing, Software

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    SouthernBiotech biotin conjugated goat anti mouse ig kappa
    In vivo expression of the IGKV3-encoded mLCV3-Tg <t>kappa</t> light chain. A. Representative flow cytometry plots of splenocytes, gated on lymphocytes by forward and side scatter and on B cells using the CD19 marker. B. Serum kappa Ig levels as determined by ELISA. Endogenous k refers to genetic sufficiency at the endogenous kappa locus. * p
    Biotin Conjugated Goat Anti Mouse Ig Kappa, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotin conjugated goat anti mouse ig kappa/product/SouthernBiotech
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotin conjugated goat anti mouse ig kappa - by Bioz Stars, 2021-07
    91/100 stars
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    99
    SouthernBiotech anti igg
    The binding of serum <t>IgG</t> to Fab fragment of IgA1 (Ste) myeloma protein. Wells of microtiter plates were coated with Fab fragment of IgA1, incubated with diluted sera from 20 IgAN patients, 20 healthy controls, and 20 patients with non-IgA GN and subsequently with <t>biotinylated</t> mAb specific for IgG, avidin-alkaline phosphatase, and phosphatase substrate. Data shown are OD at 405 nm, mean and SD. Statistical significance is noted; NS, not significant.
    Anti Igg, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti igg/product/SouthernBiotech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    93
    SouthernBiotech dylight 650 conjugated goat f ab 2 anti human λ κ
    Atypical MBCs rapidly capture and internalize PMS-bound <t>anti-λ/κ.</t> PBMCs were incubated for increasing lengths of time (30 to 120 min) at 37°C on PMS containing biotinylated anti-λ/κ conjugated to <t>DyLight</t> 650 and avidin-pHrodo. At each time point, cells were harvested and stained with streptavidin-Alexa488 and antibodies specific for CD19, CD21, and CD27 to identify naïve B cells (CD19 + CD21 + CD27 − ), classical MBCs (CD19 + CD21 + CD27 + ), and atypical MBCs (CD19 + CD21 − CD27 − ). Antigen capture, internalization, and trafficking into the acidic compartments were analyzed by flow cytometry as described in the text and detailed in Materials and Methods ( n = 4). In all cases: atypical MBCs (red circles), classical MBCs (green circles), and naïve B cells (blue circles) (see also fig. S2). ( A ) Comparison of the total amount of anti-λ/κ captured from PMS over time indicated by the MFI of DyLight 650. ( B ) Comparison of the amount of anti-λ/κ internalized in 15 min (left) and over time (right). ( C ) Comparison of anti-λ/κ trafficking to acidic compartment over time indicated by the percentage of cells positive for pHrodo or by the increases in pHrodo MFI from time 0. Data were analyzed using paired t test (A and C) or two-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (B). * P
    Dylight 650 Conjugated Goat F Ab 2 Anti Human λ κ, 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
    https://www.bioz.com/result/dylight 650 conjugated goat f ab 2 anti human λ κ/product/SouthernBiotech
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dylight 650 conjugated goat f ab 2 anti human λ κ - by Bioz Stars, 2021-07
    93/100 stars
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    94
    SouthernBiotech anti iga
    The binding of serum IgG to Fab fragment of IgA1 (Ste) myeloma protein. Wells of microtiter plates were coated with Fab fragment of IgA1, incubated with diluted sera from 20 IgAN patients, 20 healthy controls, and 20 patients with <t>non-IgA</t> GN and subsequently with <t>biotinylated</t> mAb specific for IgG, avidin-alkaline phosphatase, and phosphatase substrate. Data shown are OD at 405 nm, mean and SD. Statistical significance is noted; NS, not significant.
    Anti Iga, 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
    https://www.bioz.com/result/anti iga/product/SouthernBiotech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti iga - by Bioz Stars, 2021-07
    94/100 stars
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    Image Search Results


    In vivo expression of the IGKV3-encoded mLCV3-Tg kappa light chain. A. Representative flow cytometry plots of splenocytes, gated on lymphocytes by forward and side scatter and on B cells using the CD19 marker. B. Serum kappa Ig levels as determined by ELISA. Endogenous k refers to genetic sufficiency at the endogenous kappa locus. * p

    Journal: Molecular immunology

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities

    doi: 10.1016/j.molimm.2017.08.015

    Figure Lengend Snippet: In vivo expression of the IGKV3-encoded mLCV3-Tg kappa light chain. A. Representative flow cytometry plots of splenocytes, gated on lymphocytes by forward and side scatter and on B cells using the CD19 marker. B. Serum kappa Ig levels as determined by ELISA. Endogenous k refers to genetic sufficiency at the endogenous kappa locus. * p

    Article Snippet: Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies).

    Techniques: In Vivo, Expressing, Flow Cytometry, Cytometry, Marker, Enzyme-linked Immunosorbent Assay

    Serum mLCV3-Tg autoantibodies. Binding of serum Ig to alpha3(IV)NC1 collagen (left panel) or collagen II (right panel) was determined in ELISA. Serum was diluted 1:20 and binding detected with anti-mouse-kappa-chain Ig. The dashed line indicates the cut-off for positive binding in each assay.

    Journal: Molecular immunology

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities

    doi: 10.1016/j.molimm.2017.08.015

    Figure Lengend Snippet: Serum mLCV3-Tg autoantibodies. Binding of serum Ig to alpha3(IV)NC1 collagen (left panel) or collagen II (right panel) was determined in ELISA. Serum was diluted 1:20 and binding detected with anti-mouse-kappa-chain Ig. The dashed line indicates the cut-off for positive binding in each assay.

    Article Snippet: Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies).

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

    Spleen B cell surface kappa and lambda light chain expression. A. Percent of spleen B cells expressing surface kappa, lambda, or both kappa and lambda light chains is indicated. Values were determined by flow cytometry, after gating on lymphocytes and B cells. B. Surface kappa density on B cells expressing exclusively kappa light chains is shown. Expression is determined by mean fluorescence intensity (MFI), normalized to kappa expression on B cells of non-transgenic kappa-sufficient mice. C. Correlation between percent lambda-expressing B cells and B cell surface Ig density among B cells from mLCV3-Tg/kKO mice. Total Ig density is measured by flow cytometry as IgM+IgD MFI, normalized to spleen B cells from non-transgenic kappa-sufficient mice.

    Journal: Molecular immunology

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities

    doi: 10.1016/j.molimm.2017.08.015

    Figure Lengend Snippet: Spleen B cell surface kappa and lambda light chain expression. A. Percent of spleen B cells expressing surface kappa, lambda, or both kappa and lambda light chains is indicated. Values were determined by flow cytometry, after gating on lymphocytes and B cells. B. Surface kappa density on B cells expressing exclusively kappa light chains is shown. Expression is determined by mean fluorescence intensity (MFI), normalized to kappa expression on B cells of non-transgenic kappa-sufficient mice. C. Correlation between percent lambda-expressing B cells and B cell surface Ig density among B cells from mLCV3-Tg/kKO mice. Total Ig density is measured by flow cytometry as IgM+IgD MFI, normalized to spleen B cells from non-transgenic kappa-sufficient mice.

    Article Snippet: Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies).

    Techniques: Expressing, Flow Cytometry, Cytometry, Fluorescence, Transgenic Assay, Mouse Assay

    The binding of serum IgG to Fab fragment of IgA1 (Ste) myeloma protein. Wells of microtiter plates were coated with Fab fragment of IgA1, incubated with diluted sera from 20 IgAN patients, 20 healthy controls, and 20 patients with non-IgA GN and subsequently with biotinylated mAb specific for IgG, avidin-alkaline phosphatase, and phosphatase substrate. Data shown are OD at 405 nm, mean and SD. Statistical significance is noted; NS, not significant.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: The binding of serum IgG to Fab fragment of IgA1 (Ste) myeloma protein. Wells of microtiter plates were coated with Fab fragment of IgA1, incubated with diluted sera from 20 IgAN patients, 20 healthy controls, and 20 patients with non-IgA GN and subsequently with biotinylated mAb specific for IgG, avidin-alkaline phosphatase, and phosphatase substrate. Data shown are OD at 405 nm, mean and SD. Statistical significance is noted; NS, not significant.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Binding Assay, Incubation, Avidin-Biotin Assay

    Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques:

    Distribution of IgM, IgA, IgG, and reactivity with HAA in serum fractions from Superose 6 column. Serum from an IgAN patient was fractionated on a Superose 6 column (0.9 × 58.5 cm); 0.25-mL fractions were collected and analyzed by ELISA for the content of IgM (squares), IgA (circles), IgG (open triangles), and reactivity with HAA (filled triangles). Fractions containing aberrantly glycosylated IgA were pooled (HAA pool) and used in the inhibition experiment.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Distribution of IgM, IgA, IgG, and reactivity with HAA in serum fractions from Superose 6 column. Serum from an IgAN patient was fractionated on a Superose 6 column (0.9 × 58.5 cm); 0.25-mL fractions were collected and analyzed by ELISA for the content of IgM (squares), IgA (circles), IgG (open triangles), and reactivity with HAA (filled triangles). Fractions containing aberrantly glycosylated IgA were pooled (HAA pool) and used in the inhibition experiment.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Enzyme-linked Immunosorbent Assay, Inhibition

    Subclass specificity of serum IgG binding to a,a-IgA1. Wells of microtiter plates were coated with myeloma a,a-IgA1, incubated with sera (diluted 1:10) from 30 IgAN patients and 30 healthy controls (C) and subsequently with biotinylated mAb specific for IgG1, IgG2, IgG3, and IgG4; avidin-alkaline phosphatase; and phosphatase substrate. Data shown are OD at 405 nm, mean and SD.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Subclass specificity of serum IgG binding to a,a-IgA1. Wells of microtiter plates were coated with myeloma a,a-IgA1, incubated with sera (diluted 1:10) from 30 IgAN patients and 30 healthy controls (C) and subsequently with biotinylated mAb specific for IgG1, IgG2, IgG3, and IgG4; avidin-alkaline phosphatase; and phosphatase substrate. Data shown are OD at 405 nm, mean and SD.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Binding Assay, Incubation, Avidin-Biotin Assay

    Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques:

    Inhibition of IgA1-IgG complex formation. IgA1-IgG CICs from an IgAN patient (left columns 1–6) and a healthy control (right columns 1–6) prepared by size-exclusion chromatography on Superose 6 were dissociated at pH 3.0. Aliquots of dissociated CICs were incubated with agarose- or Sepharose-immobilized inhibitors: 1, IgA1(Mce); 2, a,a-IgA1(Mce); 3, GalNAc; 4, GlcNAc; 5, a-OSM; and 6, HSA. The mixture was adjusted to neutral pH to allow re-formation of immune complexes, and the affinity of IgG toward the particular inhibitor was determined as described in Methods.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Inhibition of IgA1-IgG complex formation. IgA1-IgG CICs from an IgAN patient (left columns 1–6) and a healthy control (right columns 1–6) prepared by size-exclusion chromatography on Superose 6 were dissociated at pH 3.0. Aliquots of dissociated CICs were incubated with agarose- or Sepharose-immobilized inhibitors: 1, IgA1(Mce); 2, a,a-IgA1(Mce); 3, GalNAc; 4, GlcNAc; 5, a-OSM; and 6, HSA. The mixture was adjusted to neutral pH to allow re-formation of immune complexes, and the affinity of IgG toward the particular inhibitor was determined as described in Methods.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Inhibition, Size-exclusion Chromatography, Incubation

    Atypical MBCs rapidly capture and internalize PMS-bound anti-λ/κ. PBMCs were incubated for increasing lengths of time (30 to 120 min) at 37°C on PMS containing biotinylated anti-λ/κ conjugated to DyLight 650 and avidin-pHrodo. At each time point, cells were harvested and stained with streptavidin-Alexa488 and antibodies specific for CD19, CD21, and CD27 to identify naïve B cells (CD19 + CD21 + CD27 − ), classical MBCs (CD19 + CD21 + CD27 + ), and atypical MBCs (CD19 + CD21 − CD27 − ). Antigen capture, internalization, and trafficking into the acidic compartments were analyzed by flow cytometry as described in the text and detailed in Materials and Methods ( n = 4). In all cases: atypical MBCs (red circles), classical MBCs (green circles), and naïve B cells (blue circles) (see also fig. S2). ( A ) Comparison of the total amount of anti-λ/κ captured from PMS over time indicated by the MFI of DyLight 650. ( B ) Comparison of the amount of anti-λ/κ internalized in 15 min (left) and over time (right). ( C ) Comparison of anti-λ/κ trafficking to acidic compartment over time indicated by the percentage of cells positive for pHrodo or by the increases in pHrodo MFI from time 0. Data were analyzed using paired t test (A and C) or two-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (B). * P

    Journal: Science Advances

    Article Title: Expression of inhibitory receptors by B cells in chronic human infectious diseases restricts responses to membrane-associated antigens

    doi: 10.1126/sciadv.aba6493

    Figure Lengend Snippet: Atypical MBCs rapidly capture and internalize PMS-bound anti-λ/κ. PBMCs were incubated for increasing lengths of time (30 to 120 min) at 37°C on PMS containing biotinylated anti-λ/κ conjugated to DyLight 650 and avidin-pHrodo. At each time point, cells were harvested and stained with streptavidin-Alexa488 and antibodies specific for CD19, CD21, and CD27 to identify naïve B cells (CD19 + CD21 + CD27 − ), classical MBCs (CD19 + CD21 + CD27 + ), and atypical MBCs (CD19 + CD21 − CD27 − ). Antigen capture, internalization, and trafficking into the acidic compartments were analyzed by flow cytometry as described in the text and detailed in Materials and Methods ( n = 4). In all cases: atypical MBCs (red circles), classical MBCs (green circles), and naïve B cells (blue circles) (see also fig. S2). ( A ) Comparison of the total amount of anti-λ/κ captured from PMS over time indicated by the MFI of DyLight 650. ( B ) Comparison of the amount of anti-λ/κ internalized in 15 min (left) and over time (right). ( C ) Comparison of anti-λ/κ trafficking to acidic compartment over time indicated by the percentage of cells positive for pHrodo or by the increases in pHrodo MFI from time 0. Data were analyzed using paired t test (A and C) or two-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (B). * P

    Article Snippet: After washing with 10 ml of 1× PMS washing buffer to remove all debris and incubating with the blocking buffer (5% BSA-containing 1× PMS buffer) for 30 min, annexin V–biotin (0.5 mg/ml) was added for binding to the PMS for 30 min and washed with 15 ml of 1× washing buffer, and this step was repeated with streptavidin (5 mg/ml) and then 10 nM biotin- and DyLight 650–conjugated goat F(ab′)2 anti-human λ + κ (SouthernBiotech).

    Techniques: Incubation, Avidin-Biotin Assay, Staining, Flow Cytometry

    The binding of serum IgG to Fab fragment of IgA1 (Ste) myeloma protein. Wells of microtiter plates were coated with Fab fragment of IgA1, incubated with diluted sera from 20 IgAN patients, 20 healthy controls, and 20 patients with non-IgA GN and subsequently with biotinylated mAb specific for IgG, avidin-alkaline phosphatase, and phosphatase substrate. Data shown are OD at 405 nm, mean and SD. Statistical significance is noted; NS, not significant.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: The binding of serum IgG to Fab fragment of IgA1 (Ste) myeloma protein. Wells of microtiter plates were coated with Fab fragment of IgA1, incubated with diluted sera from 20 IgAN patients, 20 healthy controls, and 20 patients with non-IgA GN and subsequently with biotinylated mAb specific for IgG, avidin-alkaline phosphatase, and phosphatase substrate. Data shown are OD at 405 nm, mean and SD. Statistical significance is noted; NS, not significant.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Binding Assay, Incubation, Avidin-Biotin Assay

    Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques:

    Investigations for the presence of IgA1 and IgM antibodies with anti–a,a-IgA1 binding activity.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Investigations for the presence of IgA1 and IgM antibodies with anti–a,a-IgA1 binding activity.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Binding Assay, Activity Assay

    Distribution of IgM, IgA, IgG, and reactivity with HAA in serum fractions from Superose 6 column. Serum from an IgAN patient was fractionated on a Superose 6 column (0.9 × 58.5 cm); 0.25-mL fractions were collected and analyzed by ELISA for the content of IgM (squares), IgA (circles), IgG (open triangles), and reactivity with HAA (filled triangles). Fractions containing aberrantly glycosylated IgA were pooled (HAA pool) and used in the inhibition experiment.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Distribution of IgM, IgA, IgG, and reactivity with HAA in serum fractions from Superose 6 column. Serum from an IgAN patient was fractionated on a Superose 6 column (0.9 × 58.5 cm); 0.25-mL fractions were collected and analyzed by ELISA for the content of IgM (squares), IgA (circles), IgG (open triangles), and reactivity with HAA (filled triangles). Fractions containing aberrantly glycosylated IgA were pooled (HAA pool) and used in the inhibition experiment.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Enzyme-linked Immunosorbent Assay, Inhibition

    Subclass specificity of serum IgG binding to a,a-IgA1. Wells of microtiter plates were coated with myeloma a,a-IgA1, incubated with sera (diluted 1:10) from 30 IgAN patients and 30 healthy controls (C) and subsequently with biotinylated mAb specific for IgG1, IgG2, IgG3, and IgG4; avidin-alkaline phosphatase; and phosphatase substrate. Data shown are OD at 405 nm, mean and SD.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Subclass specificity of serum IgG binding to a,a-IgA1. Wells of microtiter plates were coated with myeloma a,a-IgA1, incubated with sera (diluted 1:10) from 30 IgAN patients and 30 healthy controls (C) and subsequently with biotinylated mAb specific for IgG1, IgG2, IgG3, and IgG4; avidin-alkaline phosphatase; and phosphatase substrate. Data shown are OD at 405 nm, mean and SD.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Binding Assay, Incubation, Avidin-Biotin Assay

    Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Interactions of human serum IgG with hinge region glycans of IgA1 myeloma proteins.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques:

    Inhibition of IgA1-IgG complex formation. IgA1-IgG CICs from an IgAN patient (left columns 1–6) and a healthy control (right columns 1–6) prepared by size-exclusion chromatography on Superose 6 were dissociated at pH 3.0. Aliquots of dissociated CICs were incubated with agarose- or Sepharose-immobilized inhibitors: 1, IgA1(Mce); 2, a,a-IgA1(Mce); 3, GalNAc; 4, GlcNAc; 5, a-OSM; and 6, HSA. The mixture was adjusted to neutral pH to allow re-formation of immune complexes, and the affinity of IgG toward the particular inhibitor was determined as described in Methods.

    Journal: Journal of Clinical Investigation

    Article Title: Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies

    doi:

    Figure Lengend Snippet: Inhibition of IgA1-IgG complex formation. IgA1-IgG CICs from an IgAN patient (left columns 1–6) and a healthy control (right columns 1–6) prepared by size-exclusion chromatography on Superose 6 were dissociated at pH 3.0. Aliquots of dissociated CICs were incubated with agarose- or Sepharose-immobilized inhibitors: 1, IgA1(Mce); 2, a,a-IgA1(Mce); 3, GalNAc; 4, GlcNAc; 5, a-OSM; and 6, HSA. The mixture was adjusted to neutral pH to allow re-formation of immune complexes, and the affinity of IgG toward the particular inhibitor was determined as described in Methods.

    Article Snippet: Biotinylated polyclonal antibodies to IgM and F(ab′)2 fragment of anti-IgA and anti-IgG (all heavy chain specific) were purchased from Southern Biotechnology Associates Inc. (Birmingham, Alabama, USA).

    Techniques: Inhibition, Size-exclusion Chromatography, Incubation