【求助】有人用结晶紫染色法测定细胞增殖活性的吗?
为了便于说明,把我找到的一个crystal violet染色的protocol贴上,供大家对比讨论:
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.
Carefully remove culture medium from wells.
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
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
Wash plate 2x in tap water by immersion in a large beaker. Be careful not to lift off cells. Change tap water between washes.
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
Agitate plate on orbital shaker until color is uniform with no areas of dense coloration in bottom of wells.
Read absorbence of each well at 570 nm.
















































