Human igg4

Manufactured by Merck Group

Sourced in United States

Human IgG4 is a type of immunoglobulin G (IgG) antibody found in human serum. It plays a role in the adaptive immune response by recognizing and binding to specific antigens.

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Lab products found in correlation

Mouse igg2b clone mpc 11, by BioLegend (1 mentions) Human igm, by Merck Group (1 mentions) Human iga, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Fcr blocking reagent, by Miltenyi Biotec (1 mentions) Igg2c, by Southern Biotech (1 mentions) Igg2b, by Southern Biotech (1 mentions) Igg2a, by Southern Biotech (1 mentions)

4 protocols using human igg4

FcR Receptor Binding Assay

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Labeled antibodies: PE-conjugated anti-human FcγRI (clone 10.1, IgG1 isotype), anti-human FcγRII (clone FUN-2, IgG2b isotype), anti-humanFcγRIII (clone 3G8, IgG1 isotype), anti-human FcαRI (clone A59, IgG1 isotype), mouse IgG1 (clone MOPC-21) and mouse IgG2b (clone MPC-11) were purchased from Biolegend (San Diego, CA, USA).
Non-conjugated antibodies: anti-human FcγRI (clone 10.1, IgG1 isotype) anti-human FcγRII (clone AT10, IgG1 isotype) and anti-human FcγRIII (clone 3G8, IgG1 isotype), were obtained from Abcam (Cambridge, UK). Human IgG, human IgG1, human IgG2, human IgG3, human IgG4, human IgM, human IgA were purchased from Sigma-Aldrich, goat anti-human IgG fragment (GAH-IgG F(ab’)₂) was obtained from Jackson Immunoresearch (West Grove, PA, USA).
LPS and bovine serum albumin (BSA) were purchased from Sigma-Aldrich. FcR blocking reagent obtained from Miltenyi Biotec (Bergisch Gladbach, Germany). Viral citrullinated peptide 2 (VCP2) was prepared by solid-phase peptide synthesis and characterized by MALDI-MS at the University of Florence. Hoechst 33342 stain was from Life Technologies/Thermo Fisher (Waltham, MA, USA).

Kecse-Nagy C., Szittner Z., Papp K., Hegyi Z., Rovero P., Migliorini P., Lóránd V., Homolya L, & Prechl J. (2016). Characterization of NF-κB Reporter U937 Cells and Their Application for the Detection of Inflammatory Immune-Complexes. PLoS ONE, 11(5), e0156328.

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PD-1 Internalization with Fc Receptor Blocking

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Blood mononuclear cells were set up for surface PD-1 downregulation and internalization assay as before but in the presence of 50μg/mL of human IgG4 (Sigma) and 50μg/mL of anti-CD32/CD64 (Biolegend) for unspecific and specific blocking of the Fc receptors. Blood mononuclear cells were incubated with FcR blocking antibodies for 1 hour at 37°C before the addition of pHrodo-anti-PD-1 antibodies at 5 μg/mL and the continuation of the internalization/downregulation experiments for 24 hours and subsequent surface staining followed by flow cytometry.

Joo V., Petrovas C., de Leval L., Noto A., Obeid M., Fenwick C, & Pantaleo G. (2023). A CD64/FcγRI-mediated mechanism hijacks PD-1 from PD-L1/2 interaction and enhances anti-PD-1 functional recovery of exhausted T cells. Frontiers in Immunology, 14, 1213375.

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Neonatal Mouse Intradermal Injection Assay

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Balb/c neonatal mice (24–36 hours old, body weight 1.4–1.8 g) were intradermally injected as previously described [7 (link), 13 (link), 20 (link), 21 (link)], following IACUC UNC protocols with different quantities (contained in a volume of 100 μL) of either IgG4 or IgG1 purified fractions from FS sera. Commercially available human IgG4 (Sigma Aldrich, St Louis, MO, USA) was injected as a control. At 18–24 hours post-injection, the skin of the mice was evaluated for blisters. Skin biopsies were obtained for histology and direct immunofluorescence studies.

Evangelista F., Roth A.J., Prisayanh P., Temple B.R., Li N., Qian Y., Culton D.A., Liu Z., Harrison O.J., Brasch J., Honig B., Shapiro L, & Diaz L.A. (2018). Pathogenic IgG4 autoantibodies from endemic pemphigus foliaceus recognize a desmoglein-1 conformational epitope. Journal of autoimmunity, 89, 171-185.

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Quantification of Mouse and Human Immunoglobulin Isotypes

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Mice sera were stored at –20°C and thawed before use. We used the following capture antibodies: unlabeled goat anti-mouse IgG1 (Southern Biotech, AL, USA; 1071–01), IgG2a (Southern Biotech; 1080–01), IgG2b (Southern Biotech; 1090–01), IgG2c (Southern Biotech; 1079–01), and IgG3 (Southern Biotech; 1100–01) and rabbit anti-human IgG4 H&L (Abcam, Cambridge, UK, ab86251). These antibodies were diluted 1:100 in phosphate-buffered saline, plated, and stored at 4°C overnight. We used isotype control mouse IgG1 (Southern Biotech; 0102–01), IgG2a (Southern Biotech; 0103–01), IgG2b (Southern Biotech; 0104–01), IgG3 (Southern Biotech; 0105–01), and human IgG4 (Sigma-Aldrich, St. Louis, MO, USA; I4639) as standards. We incubated the standards and samples in 1% bovine serum albumin for 2 h. Following this, they were washed and incubated with the detection antibodies, horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG1 (Sothern Biotech; 1070–05), IgG2a (Southern Biotech; 1080–05), IgG2b (Southern Biotech; 1090–05), and IgG3 (Southern Biotech; 1100–05) at 1:5000 dilution and HRP-conjugated monoclonal HP6023 mouse anti-human IgG4 (Abcam; ab99817) at 1:2000 dilution. Subsequently, a chromogenic reagent was added to the reaction. A turbidimetric immunoassay for the quantification of human IgG4 concentration was outsourced to Japan SRL, Inc (Kumiyama, Japan).

Gon Y., Kandou T., Tsuruyama T., Iwasaki T., Kitagori K., Murakami K., Nakashima R., Akizuki S., Morinobu A., Hikida M., Mimori T, & Yoshifuji H. (2023). Increased number of T cells and exacerbated inflammatory pathophysiology in a human IgG4 knock-in MRL/lpr mouse model. PLOS ONE, 18(2), e0279389.

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