I am using different chemical like Ethanol, H2O2, toluene, ether, phenylethyl alcohol DMSO, benzene, methanol, chloroform.
Use a detergent. Saponin, digitonin, tritonx100, even tween. Mostly between 0.1-5%.
Hi Kiran, most of these reagents strip away the cell membrane (and more).
If you are looking at intracellular markers, you have to fix the cells with ~1-4% PFA or glutaraldehyde for ~20 min before you strip away the membrane with solvents.
1% - 5% Triton-X 100 is quite efficient, but be aware: it increases the risk to lose surface marker
Use a detergent. Saponin, digitonin, tritonx100, even tween. Mostly between 0.1-5%.
Kiran,
I like Daniel's choice. Saponin won't extract lpids like trition-x 100 (except for sphingolipids and cholesterol) or other solvents. It intercalates (sits between) cholesterol groups on the membranes. You must keep it in the Aby step also, or the holes will seal up. Also keep in mind that the deeper you go into the cell, the less cholesterol groups present. If you need to get to the nucleus, you will have to use Triton-x.
Hi iam doing cell cycle experiment trying several method to replace alcohol 75% so i can reduce the time. Any suggestion? Try saponin with 0.1-2% but the curve isnt clear
Please describe more detail, what do you want to do,
best regards
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