|Application Note||Recommended Starting Dilutions:|
FACS: Positive Control: Human T cells stimulated with anti-CD3 (MEM-57) antibody (1 µg/mL) Sample preparation:At the end of stimulation of T cells, perform staining of surface markers (if required) in PBS + 0.1% BSA for 20 min. on ice. Wash with PBS and fix with 2% formaldehyde, 30 min on ice. Wash with PBS and incubate in PBS + 0,1% saponine, 5 min. RT. Incubate the cells in PBS + 1% BSA + 0.03% saponine, 15 min. on ice. Incubate with fluorescence-labeled APA1/1 antibody (1-5 µg/mL) in PBS + 1% BSA + 0.03% saponine in dark, 20 min. RT.Wash with PBS + 1% BSA + 0.03% saponine, resuspend in PBS.
ICC/IF: Note:Fixed and permeabilised cells. The antibody can distinguish TCR-stimulated from non-stimulated cells
Optimal dilutions should be determined experimentally by the researcher.