Human ifnγ elisa max standard set

Manufactured by BioLegend

The Human IFNγ ELISA MAX Standard Set is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kit designed to measure human interferon-gamma (IFNγ) levels in biological samples.

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4 protocols using human ifnγ elisa max standard set

Quantifying IFNγ Levels with ELISA

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The Human IFNγ ELISA MAX Standard Set (Biolegend, London, UK) was used to quantify IFNγ concentrations in supernatants. ELISAs were performed according to the manufacturer's recommended protocol.

Houldcroft C.J., Jackson S.E., Lim E.Y., Sedikides G.X., Davies E.L., Atkinson C., McIntosh M., Remmerswaal E.B., Okecha G., Bemelman F.J., Stanton R.J., Reeves M, & Wills M.R. (2020). Assessing Anti-HCMV Cell Mediated Immune Responses in Transplant Recipients and Healthy Controls Using a Novel Functional Assay. Frontiers in Cellular and Infection Microbiology, 10, 275.

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Cytokine Secretion Assay Protocol

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Cells were pretreated with metabolic inhibitors and stimulated with antibodies or ligands as indicated above in a 24-well plate at 37°C (5% CO₂) for 4 or 6 h as indicated. The supernatant was collected and assayed for IFN-γ detection using Human IFN-γ ELISA MAX™ Standard Set (Biolegend, San Diego, CA). Granzyme B secretion was measured using Human Granzyme B ELISA development kit (Mabtech) according to the manufacturer's protocol.

Wang Z., Guan D., Wang S., Chai L.Y., Xu S, & Lam K.P. (2020). Glycolysis and Oxidative Phosphorylation Play Critical Roles in Natural Killer Cell Receptor-Mediated Natural Killer Cell Functions. Frontiers in Immunology, 11, 202.

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Quantifying Murine and Human IFNγ

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The murine IFNγ levels in culture supernatants were examined by ELISA as previously described^{114 (link)} using anti-mouse IFNγ capture and biotinylated Abs (BioLegend). The detailed information of antibodies is listed in Supplementary Table 1. The human IFNγ level in culture supernatants was examined by ELISA using the Human IFNγ ELISA MAX Standard Set (BioLegend, Cat# 430102) followed by the instructions provided by the manufacturers.

Wu M.H., Valenca-Pereira F., Cendali F., Giddings E.L., Pham-Danis C., Yarnell M.C., Novak A.J., Brunetti T.M., Thompson S.B., Henao-Mejia J., Flavell R.A., D’Alessandro A., Kohler M.E, & Rincon M. (2024). Deleting the mitochondrial respiration negative regulator MCJ enhances the efficacy of CD8+ T cell adoptive therapies in pre-clinical studies. Nature Communications, 15, 4444.

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Metabolic Modulation of Immune Cells

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Anti-CD16 antibody (clone 3G8) was purchased from BioLegend. BUV395-CD3 (clone UCHT1), PE-Cy™7-CD56 (clone B159), BV421-CD210a (clone 3F9), BV421-IgG2a (clone R35-95), FITC-CD107a (clone H4A3), PE-FasL (clone NOK-1) were purchase from BD Biosciences. Antibodies used for western blot were as follows: mouse anti-β-actin was purchased from Santa Cruz Biotechnology. Rabbit anti-phospho-S6 ribosomal protein (Ser235-Ser236) antibody, rabbit anti-phospho-p70 S6 kinase (Thr389) antibody, rabbit anti-p70 S6 kinase antibody, and mouse anti-phospho-STAT3 (Tyr705) antibody were purchased from Cell Signaling Technology.
Human IL-10 was purchased from Miltenyi Biotec, and human recombinant IL-2 was from R&D System. Cell-Tak™ Cell and Tissue Adhesive was purchased from Corning. Rapamycin and CellTrace Violet were from Invitrogen. Ficoll® Paque Plus, 2-deoxyglucose (2-DG), etomoxir, UK5099, BPTES, oligomycin, FCCP, rotenone, antimycin and concanamycin A were purchased from Sigma-Aldrich. Human IFN-γ ELISA MAX™ Standard Set was from BioLegend. Human Granzyme B ELISA development kit was from Mabtech. Lactate Assay Kit was from Sigma-Aldrich. Fixable viability dye eFluor 780 (FVD) was purchased from eBioscience. 2-NBDG was purchased from Life Technologies. Stattic was from Santa Cruz Biotechnology. SuperSignal West Pico/Dura chemiluminescent substrate was from Pierce.

Wang Z., Guan D., Huo J., Biswas S.K., Huang Y., Yang Y., Xu S, & Lam K.P. (2021). IL-10 Enhances Human Natural Killer Cell Effector Functions via Metabolic Reprogramming Regulated by mTORC1 Signaling. Frontiers in Immunology, 12, 619195.

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