Hello there.
After adding the protease inhibitor cocktail to the ripa solution, I aliquoted it at room temperature and kept at -20 degrees.
However, my aliquot process at room temperature took about 15 minutes and I did not do this on ice.
Would I damage the RIPA solution under these circumstances?
Thank all.
If you used PMSF as one of the protease inhibitors, it will be degraded because it is unstable in water. PMSF should be added fresh for each use. Everything else should be fine.
Kindly check this https://www.sigmaaldrich.com/catalog/product/sigma/r0278?lang
Prepared RIPA buffer should be aliquoted and stored at −20°C. Add protease and/or phosphatase inhibitors to a thawed aliquot before immediate use. Discard and do not freeze again. This amount is sufficient for approximately 5 million cells per 1 mL of reagent.
http://cshprotocols.cshlp.org/content/2017/12/pdb.rec101428.short
If buffer will be continually used, it is recommended that the 10x buffer be kept at 4°C for 1-2 weeks. For longer periods of time, buffer should be stored at -20°C. Aliquoting of 10x buffer is recommended if many small experiments are to be performed.
https://media.cellsignal.com/pdf/9806.pdf
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