Dear all,
I need to make a protein immunoprecipitation (IP) to find partners of the tagged protein in mammalian cell culture. I see in >90% of protocols people use frozen cell pellets.
The arguments for frozen cells are that we need an incredible amount of cells (0.2-2.0 *10^(7) cells) and nuclear fractionation without detergents, which means usage of the Dounce homogenization, and with frozen cells, nuclear extraction is more efficient.
But I worry about the complexes of interest: could it be that I destroy the complexes by freezing and thawing, and decrease the total yield of the IP?
Please, share your experience! Thank you!
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