PBMCs were resuspended at 1 × 106 cells/ml in FACS buffer with 20% human Fc receptor–binding inhibitor (eBioscience) and incubated for 20 min at room temperature in 5-ml polystyrene FACS tubes (BD Biosciences). The PBMCs were stained in a total volume of 135 μl per tube with 0.5 μl of anti-human CD3 (UCHT1) PE-Cy7 (eBioscience), 0.5 μl of anti-human CD4 (SK3) PerCP-eFluor 710 (eBioscience), 0.5 μl of anti-human CD8 (SK1) APC-eFluor780 (eBioscience), and anti-human CD14 (MφP9) V500 (BD Biosciences). The cells were incubated in the dark for 30 min at room temperature. They were washed twice with 3 ml of FACS buffer and resuspended in 0.5 ml of FACS buffer with 1 μM 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich) (fig. S11).
Anti human cd4 sk3 percp efluor 710
Manufactured by Thermo Fisher Scientific
The Anti-human CD4 (SK3) PerCP-eFluor 710 is a fluorochrome-conjugated monoclonal antibody that binds to the CD4 surface antigen expressed on human T helper cells. This product is designed for use in flow cytometry applications to identify and quantify CD4+ T cells in biological samples.
Automatically generated - may contain errors
2 protocols using anti human cd4 sk3 percp efluor 710
1
Multicolor Flow Cytometry Immunophenotyping
2
Multiplexed Intracellular Epitope Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The pooled fixed-permeabilized-barcoded cells were resuspended in 100 µl of FACS buffer containing 20% human Fc receptor binding inhibitor (eBioscience) and incubated for 20 min at room temperature in the dark. The cells were stained using 1 µl/ml anti-human CD3 (UCHT1) PE-Cy7 (eBioscience) and 1 µl/ml anti-human CD4 (SK3) PerCP-eFluor710 (eBioscience) and distributed across a 96-well polypropylene plate. The cells were stained using a Biomek NX liquid handler with fluorescently conjugated antihuman antibodies against intracellular signaling epitopes (Table S2) for 45 min in the dark at room temperature, as per the manufacturer's instructions. Antibodies against intracellular epitopes were purchased from BD Biosciences, Cell Signaling Technology and Merck Millipore and were used in groups of three antibodies per plex. The cells were washed twice and resuspended in FACS buffer at 2*10 6 cells/ml for acquisition.
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