Crystal Violet Assay 实验方法
来自 : 蚂蚁淘
CrystalVioletAssay
Thisisasimpleassayusefulforobtainingquantitativeinformationabouttherelativedensityofcellsadheringtomulti-wellclusterdishes.Thedyeinthisassay,crystalviolet,stainsDNA.Uponsolubilization,theamountofdyetakenupbythemonolayercanbequantitatedinaspectrophotometerorplatereader.
- Carefullyremoveculturemediumfromwells.
- WashplategentlywithPBSwarmedatleasttoroomtemperature:
Numberofwells | Volume |
96 | 0.2mL |
48 | 0.5mL |
24 | 1mL |
12 | 2mL |
6 | 3mL |
- CarefullyremovePBSandaddcrystalvioletsolution.Incubate10minutesatroomtemperature:
Numberofwells | Volume |
96 | 50uL |
48 | 100uL |
24 | 200uL |
12 | 500uL |
6 | 750uL |
- Washplate2xintapwaterbyimmersioninalargebeaker.Becarefulnottoliftoffcells.Changetapwaterbetweenwashes.
- Drainupsidedownonpapertowels,thanadd1%SDStosolubilizethestain:
Numberofwells | Volume |
96 | 100uL |
48 | 300uL |
24 | 600uL |
12 | 1mL |
6 | 1.5mL |
- Agitateplateonorbitalshakeruntilcolorisuniformwithnoareasofdensecolorationinbottomofwells.
- Readabsorbenceofeachwellat570nm.
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