igm from human serum  (Millipore)


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    Name:
    IgM from human serum
    Description:
    There are five immunoglobulin classes in humans Out of these immunoglobulin M IgM is a high molecular weight protein that has five or six subunits IgM monomers are made of two heavy chains and two light chains connected by a disulfide bond Human serum shows low concentration of IgM monomers It has a high carbohydrate content of about 12 It is the first immunoglobulin produced by neonates IgM antibodies are present as pentamers in the serum and are produced in response to antigens IgM from human serum is purified from normal human serum by precipitation and gel filtration techniques The immunoglobulin is determined to be atleast 95 pure by HPLC procedures
    Catalog Number:
    i8260
    Price:
    None
    Applications:
    Purified human IgM may be used as a reference antigen, standard, blocking agent, or coating protein in a variety of immunoassays including ELISA, dot immunobinding, Western immunoblotting, immunodiffusion, and immunoelectrophoresis. Other applications include starting materials for the preparation of immunogens and solid phase immunoadsorbents. IgM from human serum was used as standard in ELISA (100 μg/ml) and dot blot assay. Antibody concentration range of 200 μg/ml to 1 mg/ml was used in IgM binding assays.IgM from human serum has been used . in ELISA inhibition assay. in immunoblotting. as MW (molecular weight) standards. in IgM binding assay. to confirm the depletion of immunoglobulins
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    Structured Review

    Millipore igm from human serum
    Examples of <t>IgG1,</t> IgG2, IgG3, IgG4, <t>IgM</t> and IgE standard curves prepared with serial dilutions of the corresponding human purified isotype/subclass.
    There are five immunoglobulin classes in humans Out of these immunoglobulin M IgM is a high molecular weight protein that has five or six subunits IgM monomers are made of two heavy chains and two light chains connected by a disulfide bond Human serum shows low concentration of IgM monomers It has a high carbohydrate content of about 12 It is the first immunoglobulin produced by neonates IgM antibodies are present as pentamers in the serum and are produced in response to antigens IgM from human serum is purified from normal human serum by precipitation and gel filtration techniques The immunoglobulin is determined to be atleast 95 pure by HPLC procedures
    https://www.bioz.com/result/igm from human serum/product/Millipore
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    igm from human serum - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "Development of quantitative suspension array assays for six immunoglobulin isotypes and subclasses to multiple Plasmodium falciparum antigens"

    Article Title: Development of quantitative suspension array assays for six immunoglobulin isotypes and subclasses to multiple Plasmodium falciparum antigens

    Journal: Journal of immunological methods

    doi: 10.1016/j.jim.2018.01.009

    Examples of IgG1, IgG2, IgG3, IgG4, IgM and IgE standard curves prepared with serial dilutions of the corresponding human purified isotype/subclass.
    Figure Legend Snippet: Examples of IgG1, IgG2, IgG3, IgG4, IgM and IgE standard curves prepared with serial dilutions of the corresponding human purified isotype/subclass.

    Techniques Used: Purification

    Example of levels of antigen-specific IgG1, IgG2, IgG3, IgG4, IgM and IgE measured in plasma from European naïve individuals, at the dilutions 1:400, 1:50, 1:200, 1:50, 1:200 and 1:30, respectively.
    Figure Legend Snippet: Example of levels of antigen-specific IgG1, IgG2, IgG3, IgG4, IgM and IgE measured in plasma from European naïve individuals, at the dilutions 1:400, 1:50, 1:200, 1:50, 1:200 and 1:30, respectively.

    Techniques Used:

    2) Product Images from "Development of quantitative suspension array assays for six immunoglobulin isotypes and subclasses to multiple Plasmodium falciparum antigens"

    Article Title: Development of quantitative suspension array assays for six immunoglobulin isotypes and subclasses to multiple Plasmodium falciparum antigens

    Journal: Journal of immunological methods

    doi: 10.1016/j.jim.2018.01.009

    Examples of IgG1, IgG2, IgG3, IgG4, IgM and IgE standard curves prepared with serial dilutions of the corresponding human purified isotype/subclass.
    Figure Legend Snippet: Examples of IgG1, IgG2, IgG3, IgG4, IgM and IgE standard curves prepared with serial dilutions of the corresponding human purified isotype/subclass.

    Techniques Used: Purification

    Example of levels of antigen-specific IgG1, IgG2, IgG3, IgG4, IgM and IgE measured in plasma from European naïve individuals, at the dilutions 1:400, 1:50, 1:200, 1:50, 1:200 and 1:30, respectively.
    Figure Legend Snippet: Example of levels of antigen-specific IgG1, IgG2, IgG3, IgG4, IgM and IgE measured in plasma from European naïve individuals, at the dilutions 1:400, 1:50, 1:200, 1:50, 1:200 and 1:30, respectively.

    Techniques Used:

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Viral immunogenicity determines epidemiological fitness in a cohort of DENV-1 infection in Brazil
    Article Snippet: .. Briefly, the ELISA plates were incubated with anti-human IgM or IgG antibodies at 3 mg/mL (Sigma-Aldrich) overnight at 4°C. .. The plates were blocked with PBS containing 10% FCS for 2 h at 37°C and washed with PBS-0.05% Tween 20 (PBS-T), and serial dilutions of the supernatant samples were added and incubated overnight at 4°C.

    Incubation:

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component
    Article Snippet: .. Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C. ..

    Article Title: Intranasal Administration of a Meningococcal Outer Membrane Vesicle Vaccine Induces Persistent Local Mucosal Antibodies and Serum Antibodies with Strong Bactericidal Activity in Humans
    Article Snippet: .. After being washed with PBS containing 0.05% Tween (PBS-Tween), plates were incubated for 1 h at room temperature with horseradish peroxidase-conjugated goat antibodies specific for either human IgA, IgG, or IgM (Sigma) and developed with o -phenylenediamine (Sigma). ..

    Article Title: Viral immunogenicity determines epidemiological fitness in a cohort of DENV-1 infection in Brazil
    Article Snippet: .. Briefly, the ELISA plates were incubated with anti-human IgM or IgG antibodies at 3 mg/mL (Sigma-Aldrich) overnight at 4°C. .. The plates were blocked with PBS containing 10% FCS for 2 h at 37°C and washed with PBS-0.05% Tween 20 (PBS-T), and serial dilutions of the supernatant samples were added and incubated overnight at 4°C.

    Article Title: Isotype and IgG Subclass Distribution of Autoantibody Response to Alpha-enolase Protein in Adult Patients with Severe Asthma
    Article Snippet: .. After washes, the membrane was incubated for 2 hours with alkaline phosphatase-conjugated goat anti-human IgG, anti-human IgA, or anti-human IgM (Sigma Chemical Co., St. Louis, MO, USA) at room temperature. ..

    Article Title: Fractionation of Membrane Components from Tachyzoite Forms of Toxoplasma gondii: Differential Recognition by Immunoglobulin M (IgM) and IgG Present in Sera from Patients with Acute or Chronic Toxoplasmosis
    Article Snippet: .. Plates were then incubated with biotinylated conjugates of anti-human IgG, IgG1, IgG2, IgG3, IgG4, or IgM (Sigma) at 1:20,000 in PBS-T for additional 1 h at 37°C and washed with PBS-T. Streptavidin-peroxidase conjugate (Sigma) at a 1:1,000 dilution was added and incubated for 30 min at 37°C. ..

    other:

    Article Title: Humoral Immune Responses to Neisseria meningitidis in Children
    Article Snippet: Anti-human IgG and IgM were purchased from Sigma (Poole, United Kingdom).

    Purification:

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component
    Article Snippet: .. Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C. ..

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  • 99
    Millipore human igm
    Virus-specific pIgA for serodiagnosis of acute HEV and HAV infection. Scatterplots a show anti-HEV and anti-HAV <t>dIgA</t> compared to <t>IgM</t> reactivity detectable in acute and uninfected samples. Graph b illustrates the much higher S/Co ratio of dIgA compared to IgM in discriminating acute from uninfected samples, while graph c shows anti-HEV dIgA versus anti-HEV IgM in acute samples before and after IgM-depletion, providing evidence that the high reactivity observed for dIgA were not due to cSC cross-reactivity to IgM. Note: Unpaired and paired comparisons conducted using Mann–Whitney U and Wilcoxon test respectively, two-tailed. Asterisks indicate statistical significance of
    Human Igm, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human igm/product/Millipore
    Average 99 stars, based on 426 article reviews
    Price from $9.99 to $1999.99
    human igm - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    Virus-specific pIgA for serodiagnosis of acute HEV and HAV infection. Scatterplots a show anti-HEV and anti-HAV dIgA compared to IgM reactivity detectable in acute and uninfected samples. Graph b illustrates the much higher S/Co ratio of dIgA compared to IgM in discriminating acute from uninfected samples, while graph c shows anti-HEV dIgA versus anti-HEV IgM in acute samples before and after IgM-depletion, providing evidence that the high reactivity observed for dIgA were not due to cSC cross-reactivity to IgM. Note: Unpaired and paired comparisons conducted using Mann–Whitney U and Wilcoxon test respectively, two-tailed. Asterisks indicate statistical significance of

    Journal: BMC Research Notes

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component

    doi: 10.1186/s13104-018-3799-2

    Figure Lengend Snippet: Virus-specific pIgA for serodiagnosis of acute HEV and HAV infection. Scatterplots a show anti-HEV and anti-HAV dIgA compared to IgM reactivity detectable in acute and uninfected samples. Graph b illustrates the much higher S/Co ratio of dIgA compared to IgM in discriminating acute from uninfected samples, while graph c shows anti-HEV dIgA versus anti-HEV IgM in acute samples before and after IgM-depletion, providing evidence that the high reactivity observed for dIgA were not due to cSC cross-reactivity to IgM. Note: Unpaired and paired comparisons conducted using Mann–Whitney U and Wilcoxon test respectively, two-tailed. Asterisks indicate statistical significance of

    Article Snippet: Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C.

    Techniques: Infection, MANN-WHITNEY, Two Tailed Test

    HCV serological profile over time. a Graphs of anti-HCV antibodies compared to viral RNA or ALT over time in two seroconversion panels demonstrate that dIgA may have a different profile compared to IgA (panel 901) and it disappears even in ongoing infection (panel 400062). b Scatterplots of anti-HCV reactivity in seroconversion panels show that unlike anti-HCV IgG and IgM, anti-HCV pIgA (and to some degree anti-HCV IgA) to be statistically significantly higher in early acute phase (0–4 weeks since 1st bleed) compared to chronically infected patients. Note: Unpaired comparisons between each acute phase/early incident timepoint and chronic samples conducted using Mann–Whitney U, two tailed with Bonferonni adjustment in GraphPad Prism

    Journal: BMC Research Notes

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component

    doi: 10.1186/s13104-018-3799-2

    Figure Lengend Snippet: HCV serological profile over time. a Graphs of anti-HCV antibodies compared to viral RNA or ALT over time in two seroconversion panels demonstrate that dIgA may have a different profile compared to IgA (panel 901) and it disappears even in ongoing infection (panel 400062). b Scatterplots of anti-HCV reactivity in seroconversion panels show that unlike anti-HCV IgG and IgM, anti-HCV pIgA (and to some degree anti-HCV IgA) to be statistically significantly higher in early acute phase (0–4 weeks since 1st bleed) compared to chronically infected patients. Note: Unpaired comparisons between each acute phase/early incident timepoint and chronic samples conducted using Mann–Whitney U, two tailed with Bonferonni adjustment in GraphPad Prism

    Article Snippet: Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C.

    Techniques: Infection, MANN-WHITNEY, Two Tailed Test

    cSC preferential binding to dIgA/pIgA on ELISA and detection with monoclonal anti-human SC. Graphs show a cSC detection, b cSC-CD4 capture and c hSC-CD4 capture, to compare binding and provide dynamic range of cSC and hSC binding to human and mouse dIgA, human IgA and human IgM and d monoclonal anti-human SC antibody (AB17377; Abcam, Abingdon UK) detection of 80 kDa hSC and cSC. Note: cSC and hSC not normalized for differences in yield/concentration of active SC, error bars indicate standard error as calculated in Excel. Asterisks indicate statistical significant with reference to dIgA [p value

    Journal: BMC Research Notes

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component

    doi: 10.1186/s13104-018-3799-2

    Figure Lengend Snippet: cSC preferential binding to dIgA/pIgA on ELISA and detection with monoclonal anti-human SC. Graphs show a cSC detection, b cSC-CD4 capture and c hSC-CD4 capture, to compare binding and provide dynamic range of cSC and hSC binding to human and mouse dIgA, human IgA and human IgM and d monoclonal anti-human SC antibody (AB17377; Abcam, Abingdon UK) detection of 80 kDa hSC and cSC. Note: cSC and hSC not normalized for differences in yield/concentration of active SC, error bars indicate standard error as calculated in Excel. Asterisks indicate statistical significant with reference to dIgA [p value

    Article Snippet: Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C.

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

    Selective binding of Lmsp1 to human immunoglobulin isotypes. ELISA plates were coated with Lmsp1 (200 ng/well), blocked, and incubated in triplicate with 10 μg of purified human IgG, IgM, IgA, and IgE/ml. Detection of the isotypes bound to Lmsp1 was carried out with isotype-specific peroxidase-labeled donkey F(ab′) 2 antibody. The washing of the plates was followed by the addition of substrate (H 2 O 2 ) and chromogen (TMB). The reaction (OD) was read at 450 nm. The standard error of the triplicate OD readings was

    Journal: Infection and Immunity

    Article Title: Cloning and Characterization of a Gene Encoding an Immunoglobulin-Binding Receptor on the Cell Surface of Some Members of the Family Trypanosomatidae

    doi: 10.1128/IAI.71.9.5065-5076.2003

    Figure Lengend Snippet: Selective binding of Lmsp1 to human immunoglobulin isotypes. ELISA plates were coated with Lmsp1 (200 ng/well), blocked, and incubated in triplicate with 10 μg of purified human IgG, IgM, IgA, and IgE/ml. Detection of the isotypes bound to Lmsp1 was carried out with isotype-specific peroxidase-labeled donkey F(ab′) 2 antibody. The washing of the plates was followed by the addition of substrate (H 2 O 2 ) and chromogen (TMB). The reaction (OD) was read at 450 nm. The standard error of the triplicate OD readings was

    Article Snippet: Purified human IgG, IgA, IgM, and IgE immunoglobulin isotypes were from Calbiochem, La Jolla, Calif. Peroxidase-labeled donkey F(ab′)2 antisera specific for human, rabbit, guinea pig, goat, sheep, rat, and mouse immunoglobulins were from Zymed, South San Francisco, Calif. Rabbit anti- L. major secreted proteins and anti-purified leishmanial recombinant protein antisera were prepared as previously described ( ).

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation, Purification, Labeling

    Virus-specific pIgA for serodiagnosis of acute HEV and HAV infection. Scatterplots a show anti-HEV and anti-HAV dIgA compared to IgM reactivity detectable in acute and uninfected samples. Graph b illustrates the much higher S/Co ratio of dIgA compared to IgM in discriminating acute from uninfected samples, while graph c shows anti-HEV dIgA versus anti-HEV IgM in acute samples before and after IgM-depletion, providing evidence that the high reactivity observed for dIgA were not due to cSC cross-reactivity to IgM. Note: Unpaired and paired comparisons conducted using Mann–Whitney U and Wilcoxon test respectively, two-tailed. Asterisks indicate statistical significance of

    Journal: BMC Research Notes

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component

    doi: 10.1186/s13104-018-3799-2

    Figure Lengend Snippet: Virus-specific pIgA for serodiagnosis of acute HEV and HAV infection. Scatterplots a show anti-HEV and anti-HAV dIgA compared to IgM reactivity detectable in acute and uninfected samples. Graph b illustrates the much higher S/Co ratio of dIgA compared to IgM in discriminating acute from uninfected samples, while graph c shows anti-HEV dIgA versus anti-HEV IgM in acute samples before and after IgM-depletion, providing evidence that the high reactivity observed for dIgA were not due to cSC cross-reactivity to IgM. Note: Unpaired and paired comparisons conducted using Mann–Whitney U and Wilcoxon test respectively, two-tailed. Asterisks indicate statistical significance of

    Article Snippet: ELISA Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C.

    Techniques: Infection, MANN-WHITNEY, Two Tailed Test

    HCV serological profile over time. a Graphs of anti-HCV antibodies compared to viral RNA or ALT over time in two seroconversion panels demonstrate that dIgA may have a different profile compared to IgA (panel 901) and it disappears even in ongoing infection (panel 400062). b Scatterplots of anti-HCV reactivity in seroconversion panels show that unlike anti-HCV IgG and IgM, anti-HCV pIgA (and to some degree anti-HCV IgA) to be statistically significantly higher in early acute phase (0–4 weeks since 1st bleed) compared to chronically infected patients. Note: Unpaired comparisons between each acute phase/early incident timepoint and chronic samples conducted using Mann–Whitney U, two tailed with Bonferonni adjustment in GraphPad Prism

    Journal: BMC Research Notes

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component

    doi: 10.1186/s13104-018-3799-2

    Figure Lengend Snippet: HCV serological profile over time. a Graphs of anti-HCV antibodies compared to viral RNA or ALT over time in two seroconversion panels demonstrate that dIgA may have a different profile compared to IgA (panel 901) and it disappears even in ongoing infection (panel 400062). b Scatterplots of anti-HCV reactivity in seroconversion panels show that unlike anti-HCV IgG and IgM, anti-HCV pIgA (and to some degree anti-HCV IgA) to be statistically significantly higher in early acute phase (0–4 weeks since 1st bleed) compared to chronically infected patients. Note: Unpaired comparisons between each acute phase/early incident timepoint and chronic samples conducted using Mann–Whitney U, two tailed with Bonferonni adjustment in GraphPad Prism

    Article Snippet: ELISA Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C.

    Techniques: Infection, MANN-WHITNEY, Two Tailed Test

    cSC preferential binding to dIgA/pIgA on ELISA and detection with monoclonal anti-human SC. Graphs show a cSC detection, b cSC-CD4 capture and c hSC-CD4 capture, to compare binding and provide dynamic range of cSC and hSC binding to human and mouse dIgA, human IgA and human IgM and d monoclonal anti-human SC antibody (AB17377; Abcam, Abingdon UK) detection of 80 kDa hSC and cSC. Note: cSC and hSC not normalized for differences in yield/concentration of active SC, error bars indicate standard error as calculated in Excel. Asterisks indicate statistical significant with reference to dIgA [p value

    Journal: BMC Research Notes

    Article Title: Detection of virus-specific polymeric immunoglobulin A in acute hepatitis A, C, E virus serum samples using novel chimeric secretory component

    doi: 10.1186/s13104-018-3799-2

    Figure Lengend Snippet: cSC preferential binding to dIgA/pIgA on ELISA and detection with monoclonal anti-human SC. Graphs show a cSC detection, b cSC-CD4 capture and c hSC-CD4 capture, to compare binding and provide dynamic range of cSC and hSC binding to human and mouse dIgA, human IgA and human IgM and d monoclonal anti-human SC antibody (AB17377; Abcam, Abingdon UK) detection of 80 kDa hSC and cSC. Note: cSC and hSC not normalized for differences in yield/concentration of active SC, error bars indicate standard error as calculated in Excel. Asterisks indicate statistical significant with reference to dIgA [p value

    Article Snippet: ELISA Purified human IgA dimer (dIgA) (Nordic-MUbio; Susteren, Netherlands), mouse pIgA (in-house, 3H1-hybridoma [ ]), human IgM (Millipore; Billerica, MA), or human IgA serum standard (Nordic-MUbio; Susteren, Netherlands) at 1 μg/ml in pH9 carbonate/bicarbonate buffer diluted four-fold to 0.0625 μg/ml were incubated on 96-well Medisorp Nunc microtiter plates (Thermo Scientific; Waltham, MA) overnight at 4 °C.

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

    Binding of serum immunoglobulins and C192S. Human sera were passed through an affinity column immobilized with C192S, and then the binding ligands of C192S were eluted and the fraction 13 (F13) was collected. The contents in diluted human serum, different amount of purified human immunoglobulins, and F13 (1 µg) were verified using 12% SDS-PAGE (A) and blotted with goat anti-human IgG (B), IgM (C), or IgA (D). (E) 2 µM of purified C192S or BSA was coated in a 96-well ELISA plate, and binding of IgG, IgM, or IgA with C192S or BSA was detected by ELISA, as described in Materials and Methods. ** P

    Journal: PLoS ONE

    Article Title: Determining Antibody-Binding Site of Streptococcal Pyrogenic Exotoxin B to Protect Mice from Group A Streptococcus Infection

    doi: 10.1371/journal.pone.0055028

    Figure Lengend Snippet: Binding of serum immunoglobulins and C192S. Human sera were passed through an affinity column immobilized with C192S, and then the binding ligands of C192S were eluted and the fraction 13 (F13) was collected. The contents in diluted human serum, different amount of purified human immunoglobulins, and F13 (1 µg) were verified using 12% SDS-PAGE (A) and blotted with goat anti-human IgG (B), IgM (C), or IgA (D). (E) 2 µM of purified C192S or BSA was coated in a 96-well ELISA plate, and binding of IgG, IgM, or IgA with C192S or BSA was detected by ELISA, as described in Materials and Methods. ** P

    Article Snippet: After several washes, 50 µl of peroxidase-conjugated goat anti-human IgG, IgM, or IgA antibody (Millipore) (1∶10000) was added and incubated at 37°C for 1 h. Next, the TMB substrate (Vector Laboratories) was added, and the absorbance values were read at 650 nm.

    Techniques: Binding Assay, Affinity Column, Purification, SDS Page, Enzyme-linked Immunosorbent Assay

    Effect of SPE B on immunoglobulins. (A) Either 400 µg/ml of purified IgG or the IgM-IgA mixture was incubated with 20 µg/ml of mSPE B or C192S for 1 h with 5 mM DTT-0.1 mM EDTA. The reaction mixture was separated using 12% SDS-PAGE and blotted using goat anti-human IgG, IgM, or IgA, as described in Materials and Methods. (B) Either 400 µg/ml of purified IgG or the IgM-IgA mixture was incubated with 20 µg/ml of mSPE B for 1, 2 or 18 h with 5 mM DTT-0.1 mM EDTA. The reaction mixture was separated using 12% SDS-PAGE and blotted using goat anti-human IgG, IgM, or IgA, as described in Materials and Methods.

    Journal: PLoS ONE

    Article Title: Determining Antibody-Binding Site of Streptococcal Pyrogenic Exotoxin B to Protect Mice from Group A Streptococcus Infection

    doi: 10.1371/journal.pone.0055028

    Figure Lengend Snippet: Effect of SPE B on immunoglobulins. (A) Either 400 µg/ml of purified IgG or the IgM-IgA mixture was incubated with 20 µg/ml of mSPE B or C192S for 1 h with 5 mM DTT-0.1 mM EDTA. The reaction mixture was separated using 12% SDS-PAGE and blotted using goat anti-human IgG, IgM, or IgA, as described in Materials and Methods. (B) Either 400 µg/ml of purified IgG or the IgM-IgA mixture was incubated with 20 µg/ml of mSPE B for 1, 2 or 18 h with 5 mM DTT-0.1 mM EDTA. The reaction mixture was separated using 12% SDS-PAGE and blotted using goat anti-human IgG, IgM, or IgA, as described in Materials and Methods.

    Article Snippet: After several washes, 50 µl of peroxidase-conjugated goat anti-human IgG, IgM, or IgA antibody (Millipore) (1∶10000) was added and incubated at 37°C for 1 h. Next, the TMB substrate (Vector Laboratories) was added, and the absorbance values were read at 650 nm.

    Techniques: Purification, Incubation, SDS Page

    Binding of SPE B truncations and immunoglobulins. Either purified human IgG (A) or the IgM-IgA mixture (B) was coated in a 96-well ELISA plate, and binding of different SPE B truncations with IgG or the IgM-IgA mixture was detected by ELISA, as described in Materials and Methods. * P

    Journal: PLoS ONE

    Article Title: Determining Antibody-Binding Site of Streptococcal Pyrogenic Exotoxin B to Protect Mice from Group A Streptococcus Infection

    doi: 10.1371/journal.pone.0055028

    Figure Lengend Snippet: Binding of SPE B truncations and immunoglobulins. Either purified human IgG (A) or the IgM-IgA mixture (B) was coated in a 96-well ELISA plate, and binding of different SPE B truncations with IgG or the IgM-IgA mixture was detected by ELISA, as described in Materials and Methods. * P

    Article Snippet: After several washes, 50 µl of peroxidase-conjugated goat anti-human IgG, IgM, or IgA antibody (Millipore) (1∶10000) was added and incubated at 37°C for 1 h. Next, the TMB substrate (Vector Laboratories) was added, and the absorbance values were read at 650 nm.

    Techniques: Binding Assay, Purification, Enzyme-linked Immunosorbent Assay

    rSPE B 345–398 interfered with SPE B cleaving immunoglobulins. Either 400 µg/ml of purified human IgG or the IgM-IgA mixture was incubated with 20 µg/ml of mSPE B in the absence or presence of different concentrations of rSPE B 345–398 at 37°C for 1 h with 5 mM DTT-0.1 mM EDTA. The reaction mixtures were separated using 12% SDS-PAGE and then detected by western blotting with goat anti-human IgG, IgM, or IgA antibodies. Lane 1, purified immunoglobulins; lane 2, mSPE B-treated immunoglobulins; lane 3–5, SPE B-treated immunoglobulins incubated with different concentrations of rSPE B 345–398 ; in lane 3, 4, and 5 the molar ratio of rSPE B 345–398 /mSPE B was 5, 10, and 20, respectively.

    Journal: PLoS ONE

    Article Title: Determining Antibody-Binding Site of Streptococcal Pyrogenic Exotoxin B to Protect Mice from Group A Streptococcus Infection

    doi: 10.1371/journal.pone.0055028

    Figure Lengend Snippet: rSPE B 345–398 interfered with SPE B cleaving immunoglobulins. Either 400 µg/ml of purified human IgG or the IgM-IgA mixture was incubated with 20 µg/ml of mSPE B in the absence or presence of different concentrations of rSPE B 345–398 at 37°C for 1 h with 5 mM DTT-0.1 mM EDTA. The reaction mixtures were separated using 12% SDS-PAGE and then detected by western blotting with goat anti-human IgG, IgM, or IgA antibodies. Lane 1, purified immunoglobulins; lane 2, mSPE B-treated immunoglobulins; lane 3–5, SPE B-treated immunoglobulins incubated with different concentrations of rSPE B 345–398 ; in lane 3, 4, and 5 the molar ratio of rSPE B 345–398 /mSPE B was 5, 10, and 20, respectively.

    Article Snippet: After several washes, 50 µl of peroxidase-conjugated goat anti-human IgG, IgM, or IgA antibody (Millipore) (1∶10000) was added and incubated at 37°C for 1 h. Next, the TMB substrate (Vector Laboratories) was added, and the absorbance values were read at 650 nm.

    Techniques: Purification, Incubation, SDS Page, Western Blot