biotinylated anti-human iga Search Results


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  • 98
    Vector Laboratories biotinylated goat igg against human iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Goat Igg Against Human Iga, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 98/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated goat igg against human iga/product/Vector Laboratories
    Average 98 stars, based on 2 article reviews
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    92
    Thermo Fisher anti human iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Anti Human Iga, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti human iga/product/Thermo Fisher
    Average 92 stars, based on 64 article reviews
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    91
    SouthernBiotech biotinylated anti human iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Anti Human Iga, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 91/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 91 stars, based on 18 article reviews
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    93
    Millipore biotinylated anti human iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Anti Human Iga, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated anti human iga/product/Millipore
    Average 93 stars, based on 3 article reviews
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    85
    Jackson Immuno biotinylated rabbit anti human iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Rabbit Anti Human Iga, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated rabbit anti human iga/product/Jackson Immuno
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    94
    SouthernBiotech biotinylated iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Iga, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Perry Laboratory biotinylated anti human iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Anti Human Iga, supplied by Perry Laboratory, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher biotinylated mouse monoclonal anti human iga
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Mouse Monoclonal Anti Human Iga, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher biotinylated anti human igg iga antibody
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Anti Human Igg Iga Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Jackson Immuno biotinylated anti human iga igg igm
    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows <t>IgA</t> concentration and panel (c) the amount of <t>IgG–IgA</t> complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.
    Biotinylated Anti Human Iga Igg Igm, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 86/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson biotinylated mouse anti human iga1 iga2
    Breast milk <t>IgA</t> levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in infants with no cow’s milk allergy (CMA) and in those with either immediate-type or delayed-type CMA. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.
    Biotinylated Mouse Anti Human Iga1 Iga2, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam biotinylated antibodies anti iga
    Breast milk <t>IgA</t> levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in infants with no cow’s milk allergy (CMA) and in those with either immediate-type or delayed-type CMA. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.
    Biotinylated Antibodies Anti Iga, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 4 article reviews
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    88
    SouthernBiotech biotinylated goat anti human iga
    Breast milk <t>IgA</t> levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in infants with no cow’s milk allergy (CMA) and in those with either immediate-type or delayed-type CMA. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.
    Biotinylated Goat Anti Human Iga, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 88/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated goat anti human iga/product/SouthernBiotech
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    90
    Mab Technologies biotinylated goat anti human iga
    Breast milk <t>IgA</t> levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in infants with no cow’s milk allergy (CMA) and in those with either immediate-type or delayed-type CMA. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.
    Biotinylated Goat Anti Human Iga, supplied by Mab Technologies, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 3 article reviews
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    88
    Thermo Fisher biotinylated goat anti human iga
    Breast milk <t>IgA</t> levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in infants with no cow’s milk allergy (CMA) and in those with either immediate-type or delayed-type CMA. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.
    Biotinylated Goat Anti Human Iga, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Agilent technologies biotinylated rabbit anti human iga α chain specific antiserum
    Polymeric <t>IgA</t> molecules assemble in CHO cells. (A) Cell culture SN from clones 22, F, and 6 were concentrated by precipitation with TCA, or immunoprecipitation (IP) with anti-κ-chain antiserum/protein A Sepharose beads. After separation onto a 5–13% polyacrylamide gradient gel in SDS under nonreducing conditions, the proteins were transferred onto membranes and detected by using an <t>anti-α-chain</t> antiserum. Molecular weight markers run in parallel served to identify the various polymeric forms marked as combination of H (heavy), L (light), and J (J) chains and SC (SC). (B) Cell culture SN from clones F and 6 were concentrated with anti-κ-chain antiserum/protein A Sepharose beads and processed as in A . Blotted proteins were incubated with antisera directed against the κ-chain (lanes 1 and 2), the J chain (lanes 3 and 4), and SC (lane 5) to confirm the nature of species identified in A . Note that each specific antiserum has a distinct sensitivity, which explains the different relative intensity of the multiple molecular forms.
    Biotinylated Rabbit Anti Human Iga α Chain Specific Antiserum, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    SouthernBiotech biotinylated anti iga2
    Polymeric <t>IgA</t> molecules assemble in CHO cells. (A) Cell culture SN from clones 22, F, and 6 were concentrated by precipitation with TCA, or immunoprecipitation (IP) with anti-κ-chain antiserum/protein A Sepharose beads. After separation onto a 5–13% polyacrylamide gradient gel in SDS under nonreducing conditions, the proteins were transferred onto membranes and detected by using an <t>anti-α-chain</t> antiserum. Molecular weight markers run in parallel served to identify the various polymeric forms marked as combination of H (heavy), L (light), and J (J) chains and SC (SC). (B) Cell culture SN from clones F and 6 were concentrated with anti-κ-chain antiserum/protein A Sepharose beads and processed as in A . Blotted proteins were incubated with antisera directed against the κ-chain (lanes 1 and 2), the J chain (lanes 3 and 4), and SC (lane 5) to confirm the nature of species identified in A . Note that each specific antiserum has a distinct sensitivity, which explains the different relative intensity of the multiple molecular forms.
    Biotinylated Anti Iga2, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated anti iga2/product/SouthernBiotech
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    88
    Miltenyi Biotec anti human iga
    Influence of immunosuppression on immunoglobulin class-switching and secretion in vitro . Total and naive B cells were cultured for 7 days with or without immunosuppressive drugs (IS) in immunoglobulin (Ig)G-/IgE- [interleukin (IL)-4] and <t>IgA-</t> [transforming growth factor (TGF)-β] promoting conditions. Surface Ig expression was analysed by flow cytometry. Mean ± standard deviation (s.d.) of five different healthy donors without or with titrated doses of IS are plotted for IgE and IgA surface staining (a,b). Secreted IgE, IgA and <t>IgG</t> were measured by enzyme-linked immunosorbent assay (ELISA) in supernatant and the mean ± s.d. of five different healthy donors was plotted (c,d). Per donor, the measured values were divided by the value of the condition without IS (‘none'), to compensate for interdonor variability. Rapa = rapamycin; MethylPred = methylprednisolone; FK506 = tacrolimus; CyA = cyclosporin A.
    Anti Human Iga, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    SouthernBiotech human iga
    TG2-mediated cross-linking of BCRs. Murine A20 cells were retrovirally transduced with constructs encoding HLA-DQ2.5 and various BCRs. The TG2-specific mAb 679-14-E06 was expressed as either <t>IgA1</t> or <t>IgD</t> BCR, whereas TG2-specific 693-1-F06 and TG2-negative
    Human Iga, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 91/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows IgA concentration and panel (c) the amount of IgG–IgA complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.

    Journal: Results in Immunology

    Article Title: In vitro-generated immune complexes containing galactose-deficient IgA1 stimulate proliferation of mesangial cells

    doi: 10.1016/j.rinim.2012.08.002

    Figure Lengend Snippet: Proliferation of human mesangial cells stimulated with fractions of native or heat-inactivated cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg) was added to 160 μl of native (filled circles) or heat-inactivated (empty circles) cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells, and cellular proliferation was measured. Panel (a) shows relative cellular proliferation. Panel (b) shows IgA concentration and panel (c) the amount of IgG–IgA complexes in the collected fractions. V 0 , void volume. Results of one of the two experiments are shown.

    Article Snippet: Proteins separated by SDS-PAGE were blotted on Immobilon P (Millipore) and detected with biotinylated goat IgG against human IgA or IgG (Vector Laboratories, Burlingame, CA, USA) followed by horseradish peroxidase-conjugated NeutrAvidin (Pierce Chemical Company, Rockford, IL, USA).

    Techniques: Incubation, Concentration Assay

    Proliferation of human mesangial cells stimulated with immune complexes formed in vitro using cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg, filled circles, corresponding to final concentration of 10 μg/ml culture medium) was added to 160 μl cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells. Cellular proliferation was measured after overnight incubation. Panel (a) shows relative cellular proliferation (empty circles represent activity of cord-blood serum without any supplementation), panel (b) shows IgA concentration, and panel (c) shows the amount of IgG–IgA immune complexes in the collected fractions. V 0 , void volume.

    Journal: Results in Immunology

    Article Title: In vitro-generated immune complexes containing galactose-deficient IgA1 stimulate proliferation of mesangial cells

    doi: 10.1016/j.rinim.2012.08.002

    Figure Lengend Snippet: Proliferation of human mesangial cells stimulated with immune complexes formed in vitro using cord-blood serum supplemented with Gal-deficient IgA1 myeloma protein . Gal-deficient IgA1 (Mce) myeloma protein (80 μg, filled circles, corresponding to final concentration of 10 μg/ml culture medium) was added to 160 μl cord-blood serum. After overnight incubation at 4 °C to allow formation of immune complexes, the samples were fractionated on a calibrated Superose 6 column, and the resultant fractions were filter-sterilized and added to serum-starved human mesangial cells. Cellular proliferation was measured after overnight incubation. Panel (a) shows relative cellular proliferation (empty circles represent activity of cord-blood serum without any supplementation), panel (b) shows IgA concentration, and panel (c) shows the amount of IgG–IgA immune complexes in the collected fractions. V 0 , void volume.

    Article Snippet: Proteins separated by SDS-PAGE were blotted on Immobilon P (Millipore) and detected with biotinylated goat IgG against human IgA or IgG (Vector Laboratories, Burlingame, CA, USA) followed by horseradish peroxidase-conjugated NeutrAvidin (Pierce Chemical Company, Rockford, IL, USA).

    Techniques: In Vitro, Concentration Assay, Incubation, Activity Assay

    Breast milk IgA levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in infants with no cow’s milk allergy (CMA) and in those with either immediate-type or delayed-type CMA. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.

    Journal: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

    Article Title: Role of maternal elimination diets and human milk IgA in development of cow’s milk allergy in the infants

    doi: 10.1111/cea.12228

    Figure Lengend Snippet: Breast milk IgA levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in infants with no cow’s milk allergy (CMA) and in those with either immediate-type or delayed-type CMA. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.

    Article Snippet: After washing, biotinylated mouse anti-human IgA1/IgA2 (BD Pharmingen) at 1:1000, IgG 1:20000, IgG1 1:500, or IgG4 1:10000 was applied to the wells followed by incubation of 1 h at room temperature.

    Techniques:

    Breast milk IgA levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in mothers who were not eliminating cow’s milk (Milk) in their diet and in those with strict milk elimination diet. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.

    Journal: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

    Article Title: Role of maternal elimination diets and human milk IgA in development of cow’s milk allergy in the infants

    doi: 10.1111/cea.12228

    Figure Lengend Snippet: Breast milk IgA levels specific to casein (A) and beta-lactoglobulin (BLG) (B) in mothers who were not eliminating cow’s milk (Milk) in their diet and in those with strict milk elimination diet. On the box plots the median is represented by a horizontal line within the box representing the 25 th to 75 th percentile, whiskers show the 5 th to 95 th percentile and the square symbol is the geometric mean.

    Article Snippet: After washing, biotinylated mouse anti-human IgA1/IgA2 (BD Pharmingen) at 1:1000, IgG 1:20000, IgG1 1:500, or IgG4 1:10000 was applied to the wells followed by incubation of 1 h at room temperature.

    Techniques:

    Polymeric IgA molecules assemble in CHO cells. (A) Cell culture SN from clones 22, F, and 6 were concentrated by precipitation with TCA, or immunoprecipitation (IP) with anti-κ-chain antiserum/protein A Sepharose beads. After separation onto a 5–13% polyacrylamide gradient gel in SDS under nonreducing conditions, the proteins were transferred onto membranes and detected by using an anti-α-chain antiserum. Molecular weight markers run in parallel served to identify the various polymeric forms marked as combination of H (heavy), L (light), and J (J) chains and SC (SC). (B) Cell culture SN from clones F and 6 were concentrated with anti-κ-chain antiserum/protein A Sepharose beads and processed as in A . Blotted proteins were incubated with antisera directed against the κ-chain (lanes 1 and 2), the J chain (lanes 3 and 4), and SC (lane 5) to confirm the nature of species identified in A . Note that each specific antiserum has a distinct sensitivity, which explains the different relative intensity of the multiple molecular forms.

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

    Article Title: In vitro comparison of the antigen-binding and stability properties of the various molecular forms of IgA antibodies assembled and produced in CHO cells

    doi:

    Figure Lengend Snippet: Polymeric IgA molecules assemble in CHO cells. (A) Cell culture SN from clones 22, F, and 6 were concentrated by precipitation with TCA, or immunoprecipitation (IP) with anti-κ-chain antiserum/protein A Sepharose beads. After separation onto a 5–13% polyacrylamide gradient gel in SDS under nonreducing conditions, the proteins were transferred onto membranes and detected by using an anti-α-chain antiserum. Molecular weight markers run in parallel served to identify the various polymeric forms marked as combination of H (heavy), L (light), and J (J) chains and SC (SC). (B) Cell culture SN from clones F and 6 were concentrated with anti-κ-chain antiserum/protein A Sepharose beads and processed as in A . Blotted proteins were incubated with antisera directed against the κ-chain (lanes 1 and 2), the J chain (lanes 3 and 4), and SC (lane 5) to confirm the nature of species identified in A . Note that each specific antiserum has a distinct sensitivity, which explains the different relative intensity of the multiple molecular forms.

    Article Snippet: The membrane was probed for 1 h at 23°C by using the following antisera/antibodies: biotinylated rabbit anti-human IgA α-chain-specific antiserum (Dako), working dilution: 1:1,000; rabbit anti-human κ-chain-specific antiserum (Dako), working dilution: 1:3,000; rabbit anti-human J chain-specific antiserum , working dilution: 1:1,000; mouse anti-human SC-specific mAb (Sigma), working dilution: 1:3,000.

    Techniques: Cell Culture, Clone Assay, Immunoprecipitation, Molecular Weight, Incubation

    Antigen-binding activity and comparative digestion patterns of IgA molecular forms secreted by CHO cells. (A) Crude CHO SN were incubated in wells coated with RSV whole lysate containing the glycoprotein F antigen. The bound antibodies were detected with biotinylated antibodies to the α-chain, followed by streptavidin conjugated with HRP. The initial concentration of each antibody solution obtained from clones 22, F, and 6 was standardized to 20 μg/ml by diluting the SN with cell culture medium. Results are means of three separate experiments done in duplicates. SN of untransfected CHO cell clone P2 and wells noncoated with the antigen were used as controls. Lower dilutions of the SNs resulted in loss of linearity of the response curve. (B) ).

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

    Article Title: In vitro comparison of the antigen-binding and stability properties of the various molecular forms of IgA antibodies assembled and produced in CHO cells

    doi:

    Figure Lengend Snippet: Antigen-binding activity and comparative digestion patterns of IgA molecular forms secreted by CHO cells. (A) Crude CHO SN were incubated in wells coated with RSV whole lysate containing the glycoprotein F antigen. The bound antibodies were detected with biotinylated antibodies to the α-chain, followed by streptavidin conjugated with HRP. The initial concentration of each antibody solution obtained from clones 22, F, and 6 was standardized to 20 μg/ml by diluting the SN with cell culture medium. Results are means of three separate experiments done in duplicates. SN of untransfected CHO cell clone P2 and wells noncoated with the antigen were used as controls. Lower dilutions of the SNs resulted in loss of linearity of the response curve. (B) ).

    Article Snippet: The membrane was probed for 1 h at 23°C by using the following antisera/antibodies: biotinylated rabbit anti-human IgA α-chain-specific antiserum (Dako), working dilution: 1:1,000; rabbit anti-human κ-chain-specific antiserum (Dako), working dilution: 1:3,000; rabbit anti-human J chain-specific antiserum , working dilution: 1:1,000; mouse anti-human SC-specific mAb (Sigma), working dilution: 1:3,000.

    Techniques: Binding Assay, Activity Assay, Incubation, Concentration Assay, Clone Assay, Cell Culture

    Effect of time, FCS, and 2-mercaptoethanol on IgA expression. (A) Level of IgA production in SN of clones 22, F, and 6, expressed in μg per 1 × 10 6 cells as a function of time. IgA were measured by ELISA by using purified dIgA (clones 22 and F) and S-IgA (clone 6) as standards. (B) Effect of FCS and 2-mercaptoethanol (β-ME) on expression of S-IgA by clone 6. The cell culture SN was concentrated by immunoprecipitation (IP) with anti-κ-chain antiserum/protein A Sepharose beads, and separated by SDS/PAGE under reducing conditions. After transfer onto membranes, polypeptides were detected with antisera to the α-chain, the J chain, and SC. Molecular weight standards are indicated on the side of the gel. Asterisks indicate cross-reactive bands corresponding to the IgG heavy chain in the antiserum used for immunoprecipitation.

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

    Article Title: In vitro comparison of the antigen-binding and stability properties of the various molecular forms of IgA antibodies assembled and produced in CHO cells

    doi:

    Figure Lengend Snippet: Effect of time, FCS, and 2-mercaptoethanol on IgA expression. (A) Level of IgA production in SN of clones 22, F, and 6, expressed in μg per 1 × 10 6 cells as a function of time. IgA were measured by ELISA by using purified dIgA (clones 22 and F) and S-IgA (clone 6) as standards. (B) Effect of FCS and 2-mercaptoethanol (β-ME) on expression of S-IgA by clone 6. The cell culture SN was concentrated by immunoprecipitation (IP) with anti-κ-chain antiserum/protein A Sepharose beads, and separated by SDS/PAGE under reducing conditions. After transfer onto membranes, polypeptides were detected with antisera to the α-chain, the J chain, and SC. Molecular weight standards are indicated on the side of the gel. Asterisks indicate cross-reactive bands corresponding to the IgG heavy chain in the antiserum used for immunoprecipitation.

    Article Snippet: The membrane was probed for 1 h at 23°C by using the following antisera/antibodies: biotinylated rabbit anti-human IgA α-chain-specific antiserum (Dako), working dilution: 1:1,000; rabbit anti-human κ-chain-specific antiserum (Dako), working dilution: 1:3,000; rabbit anti-human J chain-specific antiserum , working dilution: 1:1,000; mouse anti-human SC-specific mAb (Sigma), working dilution: 1:3,000.

    Techniques: Expressing, Clone Assay, Enzyme-linked Immunosorbent Assay, Purification, Cell Culture, Immunoprecipitation, SDS Page, Molecular Weight

    Influence of immunosuppression on immunoglobulin class-switching and secretion in vitro . Total and naive B cells were cultured for 7 days with or without immunosuppressive drugs (IS) in immunoglobulin (Ig)G-/IgE- [interleukin (IL)-4] and IgA- [transforming growth factor (TGF)-β] promoting conditions. Surface Ig expression was analysed by flow cytometry. Mean ± standard deviation (s.d.) of five different healthy donors without or with titrated doses of IS are plotted for IgE and IgA surface staining (a,b). Secreted IgE, IgA and IgG were measured by enzyme-linked immunosorbent assay (ELISA) in supernatant and the mean ± s.d. of five different healthy donors was plotted (c,d). Per donor, the measured values were divided by the value of the condition without IS (‘none'), to compensate for interdonor variability. Rapa = rapamycin; MethylPred = methylprednisolone; FK506 = tacrolimus; CyA = cyclosporin A.

    Journal: Clinical and Experimental Immunology

    Article Title: Calcineurin inhibitors dampen humoral immunity by acting directly on naive B cells

    doi: 10.1111/cei.12604

    Figure Lengend Snippet: Influence of immunosuppression on immunoglobulin class-switching and secretion in vitro . Total and naive B cells were cultured for 7 days with or without immunosuppressive drugs (IS) in immunoglobulin (Ig)G-/IgE- [interleukin (IL)-4] and IgA- [transforming growth factor (TGF)-β] promoting conditions. Surface Ig expression was analysed by flow cytometry. Mean ± standard deviation (s.d.) of five different healthy donors without or with titrated doses of IS are plotted for IgE and IgA surface staining (a,b). Secreted IgE, IgA and IgG were measured by enzyme-linked immunosorbent assay (ELISA) in supernatant and the mean ± s.d. of five different healthy donors was plotted (c,d). Per donor, the measured values were divided by the value of the condition without IS (‘none'), to compensate for interdonor variability. Rapa = rapamycin; MethylPred = methylprednisolone; FK506 = tacrolimus; CyA = cyclosporin A.

    Article Snippet: The following monoclonal antibodies were used: anti-CD3, anti-CD19, anti-CD27, anti-CD38, anti-human IgD, anti-human IgG, anti-human IgE (all BD Biosciences), anti-human IgA (Miltenyi Biotech), avidin/streptavidin (eBioscience) and anti-human CD4 (Biolegend).

    Techniques: In Vitro, Cell Culture, Expressing, Flow Cytometry, Cytometry, Standard Deviation, Staining, Enzyme-linked Immunosorbent Assay

    Influence of immunosuppression on B cell division and differentiation in vitro . Total and naive B cells were cultured for 7 days with or without immunosuppressive drugs (IS) in immunoglobulin (Ig)G-/IgE- [interleukin (IL)-4] and IgA- [transforming growth factor (TGF)-β] promoting conditions. Proliferation and CD38 + plasmablast differentiation were analysed using flow cytometry. Mean ± standard deviation (s.d.) of five different healthy donors without or with titrated doses of IS are plotted for CD19 + alive B cell count (a,b), percentage of divided cells [carboxyfluorescein succinimidyl ester (CFSE)] (c,d) and percentage of CD38 + plasmablasts (e,f). Per donor, the measured values were divided by the value of the condition without IS (‘none'), to compensate for interdonor variability. Rapa = rapamycin; MethylPred = methylprednisolone; FK506 = tacrolimus; CyA = cyclosporin A.

    Journal: Clinical and Experimental Immunology

    Article Title: Calcineurin inhibitors dampen humoral immunity by acting directly on naive B cells

    doi: 10.1111/cei.12604

    Figure Lengend Snippet: Influence of immunosuppression on B cell division and differentiation in vitro . Total and naive B cells were cultured for 7 days with or without immunosuppressive drugs (IS) in immunoglobulin (Ig)G-/IgE- [interleukin (IL)-4] and IgA- [transforming growth factor (TGF)-β] promoting conditions. Proliferation and CD38 + plasmablast differentiation were analysed using flow cytometry. Mean ± standard deviation (s.d.) of five different healthy donors without or with titrated doses of IS are plotted for CD19 + alive B cell count (a,b), percentage of divided cells [carboxyfluorescein succinimidyl ester (CFSE)] (c,d) and percentage of CD38 + plasmablasts (e,f). Per donor, the measured values were divided by the value of the condition without IS (‘none'), to compensate for interdonor variability. Rapa = rapamycin; MethylPred = methylprednisolone; FK506 = tacrolimus; CyA = cyclosporin A.

    Article Snippet: The following monoclonal antibodies were used: anti-CD3, anti-CD19, anti-CD27, anti-CD38, anti-human IgD, anti-human IgG, anti-human IgE (all BD Biosciences), anti-human IgA (Miltenyi Biotech), avidin/streptavidin (eBioscience) and anti-human CD4 (Biolegend).

    Techniques: In Vitro, Cell Culture, Flow Cytometry, Cytometry, Standard Deviation, Cell Counting

    TG2-mediated cross-linking of BCRs. Murine A20 cells were retrovirally transduced with constructs encoding HLA-DQ2.5 and various BCRs. The TG2-specific mAb 679-14-E06 was expressed as either IgA1 or IgD BCR, whereas TG2-specific 693-1-F06 and TG2-negative

    Journal: The Journal of Immunology Author Choice

    Article Title: Igs as Substrates for Transglutaminase 2: Implications for Autoantibody Production in Celiac Disease

    doi: 10.4049/jimmunol.1501363

    Figure Lengend Snippet: TG2-mediated cross-linking of BCRs. Murine A20 cells were retrovirally transduced with constructs encoding HLA-DQ2.5 and various BCRs. The TG2-specific mAb 679-14-E06 was expressed as either IgA1 or IgD BCR, whereas TG2-specific 693-1-F06 and TG2-negative

    Article Snippet: After retroviral transduction, A20 cells were sorted based on positive staining with mAbs specific to HLA-DQ2.5 (clone 2.12.E11 conjugated to FITC [Sigma-Aldrich] prior to use), human IgD (PerCP-Cy5.5–conjugated IA6-2 [BD Biosciences]), or human IgA (biotinylated B3506B4 [SouthernBiotech] followed by PerCP-Cy5.5–conjugated streptavidin [BD Biosciences]) using a FACSAria flow cytometric cell sorter (BD Biosciences).

    Techniques: Transduction, Construct