This is a cached page for the URL ( To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
crystal violet assay

Crystal Violet Assay

This is a simple assay useful for obtaining quantitative information about the relative density of cells adhering to multi-well cluster dishes. The dye in this assay, crystal violet, stains DNA. Upon solubilization, the amount of dye taken up by the monolayer can be quantitated in a spectrophotometer or plate reader.

  1. Carefully remove culture medium from wells.
  2. Wash plate gently with PBS warmed at least to room temperature:
    Number of wells Volume
    96 0.2 mL
    48 0.5 mL
    24 1 mL
    12 2 mL
    6 3 mL
  3. Carefully remove PBS and add crystal violet solution. Incubate 10 minutes at room temperature:
    Number of wells Volume
    96 50 uL
    48 100 uL
    24 200 uL
    12 500 uL
    6 750 uL
  4. Wash plate 2x in tap water by immersion in a large beaker. Be careful not to lift off cells. Change tap water between washes.
  5. Drain upside down on paper towels, than add 1% SDS to solubilize the stain:
    Number of wells Volume
    96 100 uL
    48 300 uL
    24 600 uL
    12 1 mL
    6 1.5 mL
  6. Agitate plate on orbital shaker until color is uniform with no areas of dense coloration in bottom of wells.
  7. Read absorbence of each well at 570 nm.