Crystal Violet Assay
来自 : 蚂蚁淘
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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