Percp cy5.5 anti ifn γ

Manufactured by Thermo Fisher Scientific

PerCP-Cy5.5-anti-IFN-γ is a fluorescently-labeled antibody that binds to the cytokine interferon-gamma (IFN-γ). It is designed for use in flow cytometry applications to detect and quantify IFN-γ-producing cells.

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3 protocols using percp cy5.5 anti ifn γ

Isolation and Characterization of Infiltrated Lymphocytes

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The infiltrated lymphocytes were isolated as described previously [20] (link). The allografts were cut into pieces and digested with 2 mg/ml collagenase D (Worthington Bio, Lakewood, NJ) combined with 10% fecal calf serum in RPMI 1640 media for 2 h at 37°C. The suspensions were filtered with cell strainers (40 µm, BD Biosciences, San Diego, CA). The isolated cells were stained with FITC-anti-CD4, APC-Cy7-anti-CD11b, PerCP-Cy5.5-anti-IFN-γ, eFlour660-anti-IL-13, PE-anti-IL-17A, APC-anti-Foxp3, and 7-AAD (eBioscience, San Diego, CA) according to eBioscience’s Best Protocols. The cells were assessed with a fluorescence activated cell sorter (FACS) Aria II (BD Biosciences, San Diego, CA) and analyzed using FCS Express 4 Plus (De Novo Software, Los Angeles, CA).

Huang X., Ren L., Ye P., Cheng C., Wu J., Wang S., Sun Y., Liu Z., Xie A, & Xia J. (2014). Peroxisome Proliferator-Activated Receptor γ Deficiency in T Cells Accelerates Chronic Rejection by Influencing the Differentiation of CD4+ T Cells and Alternatively Activated Macrophages. PLoS ONE, 9(11), e112953.

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Multiparametric Profiling of T Cell Subsets

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eFluor660-anti–IL-21, Alexa Fluor 488–anti–IL-10, PerCP-Cy5.5-anti–IFN-γ, FITC-anti-CD45RA, PE-anti-ICOS, and anti-Tbet, and biotin–PD-1 were from eBioscience. Alexa Fluor 488–anti-GATA3, Alexa Fluor 647–anti-CXCR5, anti-pSTAT4, anti-pSTAT5, anti-pSTAT6, APC-anti-CD10, APC-Cy7-anti-CD4, BV605-anti-IgG, BV421-CD40L, BV711-anti–IL-2, PE-anti-pSTAT1, anti-RORγt, and Bcl-6, PE-Cy7-anti-CD25, and anti-CD27, PerCP-Cy5.5-anti-CD127, anti-pSTAT3, and anti-Tbet, SA-PerCpCy5.5, and IFN-γ were obtained from BD. Pacific Blue–anti-CD20 and SA-BV605 were purchased from BioLegend. Recombinant human IL-12 was purchased from R&D Systems. TGF-β, IL-1β, IL-6, IL-21, and IL-23 were obtained from PeproTech. PGE2 was purchased from Sigma-Aldrich. Human IL-4 and IL-10 were provided by R. de Waal Malefyt (DNAX Research Institute, Palo Alto, CA).

Ma C.S., Wong N., Rao G., Nguyen A., Avery D.T., Payne K., Torpy J., O’Young P., Deenick E., Bustamante J., Puel A., Okada S., Kobayashi M., Martinez-Barricarte R., Elliott M., Sebnem Kilic S., El Baghdadi J., Minegishi Y., Bousfiha A., Robertson N., Hambleton S., Arkwright P.D., French M., Blincoe A.K., Hsu P., Campbell D.E., Stormon M.O., Wong M., Adelstein S., Fulcher D.A., Cook M.C., Stepensky P., Boztug K., Beier R., Ikincioğullari A., Ziegler J.B., Gray P., Picard C., Boisson-Dupuis S., Phan T.G., Grimbacher B., Warnatz K., Holland S.M., Uzel G., Casanova J.L, & Tangye S.G. (2016). Unique and shared signaling pathways cooperate to regulate the differentiation of human CD4+ T cells into distinct effector subsets. The Journal of Experimental Medicine, 213(8), 1589-1608.

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Multi-Parameter Flow Cytometry Protocol

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eFluor660-anti-IL-21, PerCP-Cy5.5-anti-IFNγ, FITC-anti-CD45RA, biotin-PD-1 were from eBiosciences. Alexa647-anti-CXCR5 and anti-pSTAT1, APC-anti-CD10, APC-Cy7-anti-CD4, BV605-anti-IgG, PE-anti-pSTAT3 and anti-CCR6, Pe-Cy7-anti-CD25 and anti-CD27, PerCpCy5.5-anti-CD127, biotin-anti-IgA, SA-PerCpCy5.5, and recombinant IFNγ were from Becton Dickinson. BV421-anti-CXCR3, Pacific Blue-anti-CD20 and SA-BV605 were from Biolegend.

Ma C.S., Wong N., Rao G., Avery D.T., Torpy J., Hambridge T., Bustamante J., Okada S., Stoddard J.L., Deenick E.K., Pelham S.J., Payne K., Boisson-Dupuis S., Puel A., Kobayashi M., Arkwright P.D., Kilic S.S., Baghdadi J.E., Nonoyama S., Minegishi Y., Mahdaviani S.A., Mansouri D., Bousfiha A., Blincoe A.K., French M.A., Hsu P., Campbell D.E., Stormon M.O., Wong M., Adelstein S., Smart J.M., Fulcher D.A., Cook M.C., Phan T.G., Stepensky P., Boztug K., Kansu A., Ikincioğullari A., Baumann U., Beier R., Roscioli T., Ziegler J.B., Gray P., Picard C., Grimbacher B., Warnatz K., Holland S.M., Casanova J.L., Uzel G, & Tangye S.G. (2015). Monogenic mutations differentially impact the quantity and quality of T follicular helper cells in human primary immunodeficiencies. The Journal of allergy and clinical immunology, 136(4), 993-1006.e1.

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