Crystal Violet Assay for Proliferation (APS)

Crystal Violet Staining Protocol for Quantifying Proliferation

  1. Plate 25k cells in a 12 well plate or 50k in a 6-well plate
  2. Keep an empty well w/media to use as a blank
  3. Aspirate media, wash cells 1x with PBS
  4. Fix the cells with 10% buffered formalin for 30 min (use 1mL/well)
  5. Wash the cells 1x with ddH2O
  6. Add crystal violet solution and stain for 2 hrs
  7. Place cells on ice and rinse 3x with ddH2O
  8. Allow to dry for 5hrs-4wks
  9. Elute the dye with 10% glacial acetic acid (2ml) and read the OD at 600nm. Dilute and re-read over OD600 = 4.0.