Ki-67 Staining Protocol
Prepare 70% ethanol and chill at -20°C.
Prepare target cells of interest and wash 2X with PBS by centrifuge at 350xg for 5 minutes.
Discard supernatant and loosen the cell pellet by vortexing.
Add 3 ml cold 70% ethanol drop by drop to the cell pellet while vortexing.
Continue vortexing for 30 seconds and then incubate at -20°C for 1 hour.
Wash 3X with BioLegend’s Cell Staining Buffer (Cat. No. 420201) and then resuspend the cells at the concentration of 0.5-10 x 106/ml.
Mix 100 µl cell suspension with proper fluorochrome-conjugated Ki-67 antibody and incubate at room temperature in the dark for 30 minutes.
Wash 2X with BioLegend’s Cell Staining Buffer (Cat. No. 420201) and then resuspend in 0.5 ml cell staining buffer for flow cytometric analysis.
