miga  (ATCC)


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    ATCC miga
    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and <t>mIgA</t> <t>(2F11–10)</t> as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
    Miga, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Monoclonal antibodies directed against human FcRn and their applications"

    Article Title: Monoclonal antibodies directed against human FcRn and their applications

    Journal: mAbs

    doi: 10.4161/mabs.4.2.19397

    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
    Figure Legend Snippet: Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.

    Techniques Used: In Vivo, Injection, Enzyme-linked Immunosorbent Assay


    Structured Review

    SAS institute biochemistry 194 2023 15 28
    Biochemistry 194 2023 15 28, supplied by SAS institute, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    miga  (ATCC)


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    ATCC miga
    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and <t>mIgA</t> <t>(2F11–10)</t> as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
    Miga, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Monoclonal antibodies directed against human FcRn and their applications"

    Article Title: Monoclonal antibodies directed against human FcRn and their applications

    Journal: mAbs

    doi: 10.4161/mabs.4.2.19397

    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
    Figure Legend Snippet: Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.

    Techniques Used: In Vivo, Injection, Enzyme-linked Immunosorbent Assay

    miga  (ATCC)


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    Structured Review

    ATCC miga
    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and <t>mIgA</t> <t>(2F11–10)</t> as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
    Miga, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Monoclonal antibodies directed against human FcRn and their applications"

    Article Title: Monoclonal antibodies directed against human FcRn and their applications

    Journal: mAbs

    doi: 10.4161/mabs.4.2.19397

    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
    Figure Legend Snippet: Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.

    Techniques Used: In Vivo, Injection, Enzyme-linked Immunosorbent Assay


    Structured Review

    Othera Pharmaceuticals Inc white 531 71 5 83 15 6 254 47 8 194
    White 531 71 5 83 15 6 254 47 8 194, supplied by Othera Pharmaceuticals Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    white 531 71 5 83 15 6 254 47 8 194 - by Bioz Stars, 2024-07
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    anti tnp iga  (ATCC)


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    ATCC anti tnp iga
    Effects of <t>IgA</t> antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with <t>anti-TNP</t> IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
    Anti Tnp Iga, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils"

    Article Title: Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2022.881655

    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
    Figure Legend Snippet: Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.

    Techniques Used: Derivative Assay, Flow Cytometry, Expressing, Incubation, Purification, Activation Assay, Staining

    Surface IgA staining on bone marrow derived mast cells and peritoneal mast cells (A) Flow-cytometric evaluation of IgA binding to BMMCs. Representative histogram of αTNP IgA binding to BMMCs (top) and purified splenic T cells (middle), and a dose response curve of αTNP IgA binding to BMMCs. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. Purified splenic T cells incubated with or without anti-TNP IgA were stained with antibodies for CD3, CD4, and IgA and analyzed by flow cytometry. Data shown is one experiment representative of six independent experiments. (B) Immunofluorescence analysis of IgA binding to BMMCs. Representative photomicrographs (40x) of BMMCs incubated with or without 100µg/ml of αTNP IgA in HBSS for 30 minutes at 4°C. BMMCs were stained for nucleus (blue), IgA (green), or isotype control (middle), and merged (right). Scale bars: 75 µm. Images were taken using an EVOS M7000 microscope. Data shown is one experiment representative of two independent experiments. (C) Flow-cytometric evaluation of IgA binding to peritoneal mast cells. Representative flow cytometry plots of IgA staining on peritoneal cavity mast cells, identified by the presence of CD45, c-Kit, FcϵRIα. Data shown is one experiment representative of two independent experiments.
    Figure Legend Snippet: Surface IgA staining on bone marrow derived mast cells and peritoneal mast cells (A) Flow-cytometric evaluation of IgA binding to BMMCs. Representative histogram of αTNP IgA binding to BMMCs (top) and purified splenic T cells (middle), and a dose response curve of αTNP IgA binding to BMMCs. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. Purified splenic T cells incubated with or without anti-TNP IgA were stained with antibodies for CD3, CD4, and IgA and analyzed by flow cytometry. Data shown is one experiment representative of six independent experiments. (B) Immunofluorescence analysis of IgA binding to BMMCs. Representative photomicrographs (40x) of BMMCs incubated with or without 100µg/ml of αTNP IgA in HBSS for 30 minutes at 4°C. BMMCs were stained for nucleus (blue), IgA (green), or isotype control (middle), and merged (right). Scale bars: 75 µm. Images were taken using an EVOS M7000 microscope. Data shown is one experiment representative of two independent experiments. (C) Flow-cytometric evaluation of IgA binding to peritoneal mast cells. Representative flow cytometry plots of IgA staining on peritoneal cavity mast cells, identified by the presence of CD45, c-Kit, FcϵRIα. Data shown is one experiment representative of two independent experiments.

    Techniques Used: Staining, Derivative Assay, Binding Assay, Purification, Incubation, Flow Cytometry, Immunofluorescence, Microscopy

    Effects of calcium and sialic acid on IgA binding to BMMCs. (A) Effect of calcium chelation on IgA binding. Mean fluorescence intensity (MFI) of BMMCs stained with anti-IgA after incubation with anti-TNP IgA in the absence or presence of EGTA. (B) Retention of TNP-OVA binding by desialylated IgA. Serial dilutions of desialylated or buffer control treated anti-TNP IgA were tested for TNP-OVA binding by ELISA. (C) Analysis of sialic acid requirement for IgA binding . MFI of BMMCs incubated with untreated anti-TNP IgA, desialylated anti-TNP IgA or N-deglycosylated anti-TNP IgA in HBSS for 30 minutes at 4°C. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. (D) Effects of sialic acid competition on IgA binding to BMMCs. MFI of BMMCs stained with anti-IgA with or without preincubation with sialic acid (E) Consequences of sialic acid removal on the inhibitory effects of IgA . Bar plot of percent degranulation across three replicates of LAMP-1 induction in IgE sensitized BMMCs incubated with or without anti-TNP IgA, or desialylated anti-TNP (desial αTNP) IgA as in Figure 3A. Statistical analysis done by one-way analysis of variance (ANOVA). Data shown mean ± SEM of one experiment representative of three independent experiments. *P < .05, and ****P < .0001.
    Figure Legend Snippet: Effects of calcium and sialic acid on IgA binding to BMMCs. (A) Effect of calcium chelation on IgA binding. Mean fluorescence intensity (MFI) of BMMCs stained with anti-IgA after incubation with anti-TNP IgA in the absence or presence of EGTA. (B) Retention of TNP-OVA binding by desialylated IgA. Serial dilutions of desialylated or buffer control treated anti-TNP IgA were tested for TNP-OVA binding by ELISA. (C) Analysis of sialic acid requirement for IgA binding . MFI of BMMCs incubated with untreated anti-TNP IgA, desialylated anti-TNP IgA or N-deglycosylated anti-TNP IgA in HBSS for 30 minutes at 4°C. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. (D) Effects of sialic acid competition on IgA binding to BMMCs. MFI of BMMCs stained with anti-IgA with or without preincubation with sialic acid (E) Consequences of sialic acid removal on the inhibitory effects of IgA . Bar plot of percent degranulation across three replicates of LAMP-1 induction in IgE sensitized BMMCs incubated with or without anti-TNP IgA, or desialylated anti-TNP (desial αTNP) IgA as in Figure 3A. Statistical analysis done by one-way analysis of variance (ANOVA). Data shown mean ± SEM of one experiment representative of three independent experiments. *P < .05, and ****P < .0001.

    Techniques Used: Binding Assay, Fluorescence, Staining, Incubation, Enzyme-linked Immunosorbent Assay, Flow Cytometry

    Effect of IgA antibodies on IgE-induced phosphorylation of Syk and on cytokine production by activated bone marrow derived mast cells (A) IgA effects on Syk phosphorylation . IgE sensitized BMMCs were incubated with or without anti-TNP IgA and stimulated with antigen for up to two minutes followed by measurement of phosphorylated-Syk (phospho-Syk) using flow cytometry (upper panel). Representative Syk phosphorylation blots and compiled ratios of phospho protein/total protein intensities in anti-TNP IgE-sensitized BMMCs or anti-TNP IgE/IgA treated BMMCs (lower panel). This experiment is representative of three replicates. (B) IgA effects on cytokine production . Cytokine (IL-6, IL-13 and TNF-α) levels in the supernatants of anti-TNP IgE sensitized BMMC incubated with or without anti-TNP IgA and stimulated with TNP-BSA for 6 hours. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ***P < .001.
    Figure Legend Snippet: Effect of IgA antibodies on IgE-induced phosphorylation of Syk and on cytokine production by activated bone marrow derived mast cells (A) IgA effects on Syk phosphorylation . IgE sensitized BMMCs were incubated with or without anti-TNP IgA and stimulated with antigen for up to two minutes followed by measurement of phosphorylated-Syk (phospho-Syk) using flow cytometry (upper panel). Representative Syk phosphorylation blots and compiled ratios of phospho protein/total protein intensities in anti-TNP IgE-sensitized BMMCs or anti-TNP IgE/IgA treated BMMCs (lower panel). This experiment is representative of three replicates. (B) IgA effects on cytokine production . Cytokine (IL-6, IL-13 and TNF-α) levels in the supernatants of anti-TNP IgE sensitized BMMC incubated with or without anti-TNP IgA and stimulated with TNP-BSA for 6 hours. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ***P < .001.

    Techniques Used: Derivative Assay, Incubation, Flow Cytometry

    frozen tib 194 hybridoma cells  (ATCC)


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    Structured Review

    ATCC frozen tib 194 hybridoma cells
    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from <t>TIB-194</t> hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
    Frozen Tib 194 Hybridoma Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils"

    Article Title: Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2022.881655

    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
    Figure Legend Snippet: Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.

    Techniques Used: Derivative Assay, Flow Cytometry, Expressing, Incubation, Purification, Activation Assay, Staining


    Structured Review

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    WILEY-VCH Verlag cr 194 15 grape seed activated carbon
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    tfse 6 stne 15 tdfd 19 svve 109 ttsd 194 sene 434  (ATUM Bio)

     
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    ATUM Bio tfse 6 stne 15 tdfd 19 svve 109 ttsd 194 sene 434
    Location and number of types of post-translational modifications of octopamine receptors in Aethina tumida . The residues involved in post-translational modifications is shown below and the amino acid position in the peptide of the first residue in the motif is shown as a number
    Tfse 6 Stne 15 Tdfd 19 Svve 109 Ttsd 194 Sene 434, supplied by ATUM Bio, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "In silico identification and assessment of insecticide target sites in the genome of the small hive beetle, Aethina tumida"

    Article Title: In silico identification and assessment of insecticide target sites in the genome of the small hive beetle, Aethina tumida

    Journal: BMC Genomics

    doi: 10.1186/s12864-020-6551-y

    Location and number of types of post-translational modifications of octopamine receptors in Aethina tumida . The residues involved in post-translational modifications is shown below and the amino acid position in the peptide of the first residue in the motif is shown as a number
    Figure Legend Snippet: Location and number of types of post-translational modifications of octopamine receptors in Aethina tumida . The residues involved in post-translational modifications is shown below and the amino acid position in the peptide of the first residue in the motif is shown as a number

    Techniques Used:

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    ATCC miga
    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and <t>mIgA</t> <t>(2F11–10)</t> as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
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    SAS institute biochemistry 194 2023 15 28
    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and <t>mIgA</t> <t>(2F11–10)</t> as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
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    Othera Pharmaceuticals Inc white 531 71 5 83 15 6 254 47 8 194
    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and <t>mIgA</t> <t>(2F11–10)</t> as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.
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    ATCC anti tnp iga
    Effects of <t>IgA</t> antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with <t>anti-TNP</t> IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
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    ATCC frozen tib 194 hybridoma cells
    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from <t>TIB-194</t> hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
    Frozen Tib 194 Hybridoma Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MicroPort Scientific Corporation 194 29 101 15 medtronic
    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from <t>TIB-194</t> hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
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    WILEY-VCH Verlag cr 194 15 grape seed activated carbon
    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from <t>TIB-194</t> hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
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    VELP Scientifica 15 194 2021 temperatures
    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from <t>TIB-194</t> hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.
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    ATUM Bio tfse 6 stne 15 tdfd 19 svve 109 ttsd 194 sene 434
    Location and number of types of post-translational modifications of octopamine receptors in Aethina tumida . The residues involved in post-translational modifications is shown below and the amino acid position in the peptide of the first residue in the motif is shown as a number
    Tfse 6 Stne 15 Tdfd 19 Svve 109 Ttsd 194 Sene 434, supplied by ATUM Bio, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.

    Journal: mAbs

    Article Title: Monoclonal antibodies directed against human FcRn and their applications

    doi: 10.4161/mabs.4.2.19397

    Figure Lengend Snippet: Blockade of IgG by anti-hFcRn mAbs in vivo. (A) B6 Tg276 mice were co-injected ip with 100 µg each of hIgG, mIgG1 (1B7.11) and mIgA (2F11–10) as tracers. The mice were then injected with 1,000 µg (◊), 300 µg (◆), 100 µg (△), 30 µg (▴) or 0 µg (□) of DVN24 or with 1,000 µg (X) of a control IgG2a mAb into groups of 3 mice on days 2, 3 and 4 d after tracer injection. (B) B6 Tg276 mice (n = 3) were co-injected ip with 100 µg hIgG and 1 mg of HSA tracers followed by injections with 1 mg doses of ADM32 (−), DVN24 (◊) or isotype matched mIgG2a (X). (C) B6 Tg276 mice (n = 4–5) were injected with 100 µg hIgG tracer and then treated on days 5–7 or 10–12 with 1 mg of DVN24 (◊) or mIgG2a (X). Significant differences (A–C) are indicated as *p < 0.05 and **p < 0.01 comparing 1,000 µg injections of DVN24 with the control isotype matched mIgG2a. (D) B6 WT mice (n = 8) were injected ip with 200 µg of mIgG1 (1B7.11) as a tracer, and then injected ip on days 4–6 with 1 mg doses of DVN24 (◊), ADM31 (◆) or vehicle only (X). Tracer plasma concentrations were determined by ELISA and plotted either as percent remaining as compared with the first time point plasma concentrations or as plasma concentrations ±SD. Significant differences (D) are indicated as **p < 0.01 comparing 1,000 µg injections of DVN24 with the vehicle only control.

    Article Snippet: hFcRn Tg276 mice carrying one copy of the hFcRn or B6 WT mice were first injected i.p. with 100 µg of tracer mIgG 1 (1B7-11, anti-TNP, American Type Culture Collection, TIB-191), mIgA (2F11-15, anti-TNP, American Type Culture Collection, TIB-194), human IgG (Gammagard, Baxter, 1501375) or 1 mg HSA (Sigma-Aldrich, A-7284) as tracers.

    Techniques: In Vivo, Injection, Enzyme-linked Immunosorbent Assay

    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.

    Journal: Frontiers in Immunology

    Article Title: Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils

    doi: 10.3389/fimmu.2022.881655

    Figure Lengend Snippet: Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.

    Article Snippet: For the lysosomal-associated membrane protein 1 (LAMP-1) assay, mouse BMMCs were incubated overnight with anti-TNP IgE (50 ng/ml, BD Biosciences, Franklin Lakes, NJ), and anti-TNP IgA (100 μg/ml, TIB-194 hybridoma, ATCC, Manassas, VA).

    Techniques: Derivative Assay, Flow Cytometry, Expressing, Incubation, Purification, Activation Assay, Staining

    Surface IgA staining on bone marrow derived mast cells and peritoneal mast cells (A) Flow-cytometric evaluation of IgA binding to BMMCs. Representative histogram of αTNP IgA binding to BMMCs (top) and purified splenic T cells (middle), and a dose response curve of αTNP IgA binding to BMMCs. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. Purified splenic T cells incubated with or without anti-TNP IgA were stained with antibodies for CD3, CD4, and IgA and analyzed by flow cytometry. Data shown is one experiment representative of six independent experiments. (B) Immunofluorescence analysis of IgA binding to BMMCs. Representative photomicrographs (40x) of BMMCs incubated with or without 100µg/ml of αTNP IgA in HBSS for 30 minutes at 4°C. BMMCs were stained for nucleus (blue), IgA (green), or isotype control (middle), and merged (right). Scale bars: 75 µm. Images were taken using an EVOS M7000 microscope. Data shown is one experiment representative of two independent experiments. (C) Flow-cytometric evaluation of IgA binding to peritoneal mast cells. Representative flow cytometry plots of IgA staining on peritoneal cavity mast cells, identified by the presence of CD45, c-Kit, FcϵRIα. Data shown is one experiment representative of two independent experiments.

    Journal: Frontiers in Immunology

    Article Title: Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils

    doi: 10.3389/fimmu.2022.881655

    Figure Lengend Snippet: Surface IgA staining on bone marrow derived mast cells and peritoneal mast cells (A) Flow-cytometric evaluation of IgA binding to BMMCs. Representative histogram of αTNP IgA binding to BMMCs (top) and purified splenic T cells (middle), and a dose response curve of αTNP IgA binding to BMMCs. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. Purified splenic T cells incubated with or without anti-TNP IgA were stained with antibodies for CD3, CD4, and IgA and analyzed by flow cytometry. Data shown is one experiment representative of six independent experiments. (B) Immunofluorescence analysis of IgA binding to BMMCs. Representative photomicrographs (40x) of BMMCs incubated with or without 100µg/ml of αTNP IgA in HBSS for 30 minutes at 4°C. BMMCs were stained for nucleus (blue), IgA (green), or isotype control (middle), and merged (right). Scale bars: 75 µm. Images were taken using an EVOS M7000 microscope. Data shown is one experiment representative of two independent experiments. (C) Flow-cytometric evaluation of IgA binding to peritoneal mast cells. Representative flow cytometry plots of IgA staining on peritoneal cavity mast cells, identified by the presence of CD45, c-Kit, FcϵRIα. Data shown is one experiment representative of two independent experiments.

    Article Snippet: For the lysosomal-associated membrane protein 1 (LAMP-1) assay, mouse BMMCs were incubated overnight with anti-TNP IgE (50 ng/ml, BD Biosciences, Franklin Lakes, NJ), and anti-TNP IgA (100 μg/ml, TIB-194 hybridoma, ATCC, Manassas, VA).

    Techniques: Staining, Derivative Assay, Binding Assay, Purification, Incubation, Flow Cytometry, Immunofluorescence, Microscopy

    Effects of calcium and sialic acid on IgA binding to BMMCs. (A) Effect of calcium chelation on IgA binding. Mean fluorescence intensity (MFI) of BMMCs stained with anti-IgA after incubation with anti-TNP IgA in the absence or presence of EGTA. (B) Retention of TNP-OVA binding by desialylated IgA. Serial dilutions of desialylated or buffer control treated anti-TNP IgA were tested for TNP-OVA binding by ELISA. (C) Analysis of sialic acid requirement for IgA binding . MFI of BMMCs incubated with untreated anti-TNP IgA, desialylated anti-TNP IgA or N-deglycosylated anti-TNP IgA in HBSS for 30 minutes at 4°C. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. (D) Effects of sialic acid competition on IgA binding to BMMCs. MFI of BMMCs stained with anti-IgA with or without preincubation with sialic acid (E) Consequences of sialic acid removal on the inhibitory effects of IgA . Bar plot of percent degranulation across three replicates of LAMP-1 induction in IgE sensitized BMMCs incubated with or without anti-TNP IgA, or desialylated anti-TNP (desial αTNP) IgA as in Figure 3A. Statistical analysis done by one-way analysis of variance (ANOVA). Data shown mean ± SEM of one experiment representative of three independent experiments. *P < .05, and ****P < .0001.

    Journal: Frontiers in Immunology

    Article Title: Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils

    doi: 10.3389/fimmu.2022.881655

    Figure Lengend Snippet: Effects of calcium and sialic acid on IgA binding to BMMCs. (A) Effect of calcium chelation on IgA binding. Mean fluorescence intensity (MFI) of BMMCs stained with anti-IgA after incubation with anti-TNP IgA in the absence or presence of EGTA. (B) Retention of TNP-OVA binding by desialylated IgA. Serial dilutions of desialylated or buffer control treated anti-TNP IgA were tested for TNP-OVA binding by ELISA. (C) Analysis of sialic acid requirement for IgA binding . MFI of BMMCs incubated with untreated anti-TNP IgA, desialylated anti-TNP IgA or N-deglycosylated anti-TNP IgA in HBSS for 30 minutes at 4°C. Mast cells incubated with or without anti-TNP IgA were stained with antibodies for c-Kit, FcϵRIα, and IgA and analyzed by flow cytometry. (D) Effects of sialic acid competition on IgA binding to BMMCs. MFI of BMMCs stained with anti-IgA with or without preincubation with sialic acid (E) Consequences of sialic acid removal on the inhibitory effects of IgA . Bar plot of percent degranulation across three replicates of LAMP-1 induction in IgE sensitized BMMCs incubated with or without anti-TNP IgA, or desialylated anti-TNP (desial αTNP) IgA as in Figure 3A. Statistical analysis done by one-way analysis of variance (ANOVA). Data shown mean ± SEM of one experiment representative of three independent experiments. *P < .05, and ****P < .0001.

    Article Snippet: For the lysosomal-associated membrane protein 1 (LAMP-1) assay, mouse BMMCs were incubated overnight with anti-TNP IgE (50 ng/ml, BD Biosciences, Franklin Lakes, NJ), and anti-TNP IgA (100 μg/ml, TIB-194 hybridoma, ATCC, Manassas, VA).

    Techniques: Binding Assay, Fluorescence, Staining, Incubation, Enzyme-linked Immunosorbent Assay, Flow Cytometry

    Effect of IgA antibodies on IgE-induced phosphorylation of Syk and on cytokine production by activated bone marrow derived mast cells (A) IgA effects on Syk phosphorylation . IgE sensitized BMMCs were incubated with or without anti-TNP IgA and stimulated with antigen for up to two minutes followed by measurement of phosphorylated-Syk (phospho-Syk) using flow cytometry (upper panel). Representative Syk phosphorylation blots and compiled ratios of phospho protein/total protein intensities in anti-TNP IgE-sensitized BMMCs or anti-TNP IgE/IgA treated BMMCs (lower panel). This experiment is representative of three replicates. (B) IgA effects on cytokine production . Cytokine (IL-6, IL-13 and TNF-α) levels in the supernatants of anti-TNP IgE sensitized BMMC incubated with or without anti-TNP IgA and stimulated with TNP-BSA for 6 hours. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ***P < .001.

    Journal: Frontiers in Immunology

    Article Title: Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils

    doi: 10.3389/fimmu.2022.881655

    Figure Lengend Snippet: Effect of IgA antibodies on IgE-induced phosphorylation of Syk and on cytokine production by activated bone marrow derived mast cells (A) IgA effects on Syk phosphorylation . IgE sensitized BMMCs were incubated with or without anti-TNP IgA and stimulated with antigen for up to two minutes followed by measurement of phosphorylated-Syk (phospho-Syk) using flow cytometry (upper panel). Representative Syk phosphorylation blots and compiled ratios of phospho protein/total protein intensities in anti-TNP IgE-sensitized BMMCs or anti-TNP IgE/IgA treated BMMCs (lower panel). This experiment is representative of three replicates. (B) IgA effects on cytokine production . Cytokine (IL-6, IL-13 and TNF-α) levels in the supernatants of anti-TNP IgE sensitized BMMC incubated with or without anti-TNP IgA and stimulated with TNP-BSA for 6 hours. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ***P < .001.

    Article Snippet: For the lysosomal-associated membrane protein 1 (LAMP-1) assay, mouse BMMCs were incubated overnight with anti-TNP IgE (50 ng/ml, BD Biosciences, Franklin Lakes, NJ), and anti-TNP IgA (100 μg/ml, TIB-194 hybridoma, ATCC, Manassas, VA).

    Techniques: Derivative Assay, Incubation, Flow Cytometry

    Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.

    Journal: Frontiers in Immunology

    Article Title: Allergen-Specific IgA Antibodies Block IgE-Mediated Activation of Mast Cells and Basophils

    doi: 10.3389/fimmu.2022.881655

    Figure Lengend Snippet: Effects of IgA antibodies on IgE mediated degranulation of bone marrow derived mast cells. Flow cytometry plots from a representative experiment (left) and aggregate data (n=3) bar plots (right) of percent LAMP-1 expression of IgE sensitized BMMCs following antigen exposure. BMMCs were sensitized with anti-TNP IgE (αTNP IgE) (50 ng/ml). Subsequently, some cells were co-incubated with anti-TNP IgA (αTNP IgA) (100 µg/ml) (purified from TIB-194 hybridoma) or anti-OVA IgA [αOVA IgA (100 µg/ml)]. Cells were primed with antibodies overnight, then washed and stimulated with, 50 ng/ml of TNP-BSA for 10 minutes before assessing activation by staining with anti-mouse c-Kit, and LAMP-1. Statistical analysis done by ANOVA. Data shown mean ± SEM of one experiment representative of three independent experiments. ****P < .0001.

    Article Snippet: Frozen TIB-194 hybridoma cells (ATCC, Manassas, VA) were thawed according to manufacturer’s instructions, transferred and diluted with warmed RPMI-1640 (10% FBS, 100U/ml penicillin).

    Techniques: Derivative Assay, Flow Cytometry, Expressing, Incubation, Purification, Activation Assay, Staining

    Location and number of types of post-translational modifications of octopamine receptors in Aethina tumida . The residues involved in post-translational modifications is shown below and the amino acid position in the peptide of the first residue in the motif is shown as a number

    Journal: BMC Genomics

    Article Title: In silico identification and assessment of insecticide target sites in the genome of the small hive beetle, Aethina tumida

    doi: 10.1186/s12864-020-6551-y

    Figure Lengend Snippet: Location and number of types of post-translational modifications of octopamine receptors in Aethina tumida . The residues involved in post-translational modifications is shown below and the amino acid position in the peptide of the first residue in the motif is shown as a number

    Article Snippet: Atum_Octβ1R , NETD 17 NNTS 26 NTSI 27 NFSV 107 NRTY 214 NYSN 402 NASS 405 NISE 432 , TFSE 6 STNE 15 TDFD 19 SVVE 109 TTSD 194 SENE 434 , SLR 276 SSK 296 TSK 385 , GISAGL 269 GIIVSA 310 , , .

    Techniques: