BioLegend
PE anti-human CD279 (PD-1) Antibody
329906
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One of the interests of our lab is T-cell exhaustion in the context of viral infection and cancer. Quality reagents are required to study T-cell exhaustion cell markers by flow cytometry given their relatively low baseline expression under normal conditions as well as the relatively low frequency of expressing cells under physiological conditions. The purpose of this experiment was to establish an in vitro assay to measure the expression of T cell exhaustion markers. This particular antibody was chosen since it is (a) directly labelled with a fluorophore, (b) already titrated for use in flow cytometry, and (c) given BioLegend's exceptional reputation.
Flow Cytometry
Human peripheral blood mononuclear cells stimulated overnight with 25ng/ml phorbol 12-myristate 13-acetate and 100ng/ml ionomycin
Approximately 1x10^6 cells were incubated on ice for 1 hour in the dark with PE anti-human CD279 (PD-1) Antibody (329906; clone EH12.2H7) at a 1:20 dilution in a total volume of 100ul PBS+2% FCS, 1mM EDTA. At the same time, the cells were stained with a mouse-anti-human CD3 antibody.
N/A
BD FACSCalibur
PD-1 expression was expressed primarily on CD3+ T cells after T-cell stimulation with PMA/Ionomcyin as expected. 25.8% of CD3+ cells expressed PD-1 after overnight stimulation with PMA/Ionomycin compared to 8.5% in unstimulated control cells. Compared to a matched isotype control, PD-1 staining was distinct and easily visualized.
Directly labelled with a fluorophore. Comes titrated so minimal optimization.
None
This is a very good antibody for analysis of PD-1 expression on human blood cells by flow cytometry. Highly recommended.