I have read that MG132 should be pretreated , while few treated together. Which option will be better?
Pretreatment of MG132 for 1-2 h followed by stimulant treatment, is the experimental scheme that I follow in routine. In most of the cases I use leukemic cells.
Thank you.
How do you dilute your MG132 . I have dissoved in DMSO at 100mM. But when I diluted it with PBS, it formed precipitates.
DMSO is fine but I would suggest you to decrease the stock concentration to 20mM or at the most 50mM. Because the solubility of MG132 in DMSO is not that much high.
make 10 mM, stock and it is toxic to cell, Cells don't like it for long time. Pretreat cell for 4-6 hours and give your treatment.
Parash....DMSO induces differentiation in leukemic cells and have effects in other cells also....As per literature its final concentration given to the cells should be less than 0.1%....If we prepare a low concentration stock then it may increase the volume of DMSO treated to the cells....So I think if we keep the stock concentration a little higher that would be better....
Yes Imran, I forgot to mention that, we also keep DMSO concentration less than 0.1%, High concentration is toxic to cells.
Monika: what is the final concentration you used to treat the cell? I read in one website, they recommend me use from 1-10uM. Is this concentration you are using to treat cell?Thanks
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