Ki-67 Flow Cytometry Staining Protocol
- Prepare 70% Ethanol and chill to -20°C
Tip: Do not freeze ethanol for long-term storage.
- Prepare target cells of interest and wash 2X with PBS, centrifuging at 350xg for 5 minutes.
- Discard supernatant and loosen the cell pellet by vortexing.
- Add 3ml 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 100µl 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 and then resuspend in 0.5ml cell staining buffer for fluorescence activated cell sorting (FACS), or flow cytometric analysis.
Tip: Based on customer testing, Ki-67 staining is not recommended with our True-Nuclear™ Transcription Factor Buffer Set.