PBMCs were isolated from whole blood by Ficoll-Paque density gradient centrifugation (GE Healthcare Bio-sciences), as described previously (55 (link)). CD8+ T cells were subsequently isolated from PBMCs by negative selection using the EasySep Human CD8+ T Cell Enrichment Kit (STEMCELL Technologies Inc.) according to the manufacturer’s instructions. The CD8+ T cells were maintained in RPMI-1640 medium supplemented with 10% human serum, 200 u/ml penicillin, 200 µg/ml streptomycin, 1 mM L-glutamine, and 10 mM HEPES (55 (link)). Cells were activated for 72–96 h in a cell culture dish coated with 10 µg/ml mouse anti-human CD3 antibody (Biolegend) and 10 µg/ml mouse anti-human CD28 antibody (Biolegend).
Mouse anti human cd3 antibody
Manufactured by BioLegend
The Mouse anti-human CD3 antibody is a primary antibody that binds specifically to the CD3 complex on the surface of human T cells. The CD3 complex is essential for T cell activation and signal transduction. This antibody can be used for the identification and enumeration of T cells in flow cytometry applications.
Automatically generated - may contain errors
2 protocols using mouse anti human cd3 antibody
1
CD8+ T Cell Isolation and Activation
2
PBMC Isolation and CD8+ T Cell Activation
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Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by Ficoll-Paque density gradient centrifugation (GE Healthcare Bio-Sciences), as described previously (Chimote et al., 2018 (link)). CD8+ T cells were isolated from PBMCs by negative selection using the EasySep Human CD8+ T Cell Enrichment Kit (STEMCELL Technologies Inc.). The CD8+ T cells were maintained in RPMI 1640 medium supplemented with 10% human serum, penicillin (200 U/ml), streptomycin (200 μg/ml), 1 mM L-glutamine, and 10 mM Hepes. For all experiments, cells were activated in a cell culture dish pre-coated with mouse anti-human CD3 antibody (10 μg/ml) (BioLegend) and mouse anti-human CD28 antibody (10 μg/ml) (BioLegend) for 72 to 96 h, except where specified.
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