Which method is suitable for precipitating protein from yeast cell lysate? From my own search, I found that the method of precipitating the protein is not a single method because there are a few possible methods such as TCA/acetone, methanol/chloroform and ammonium sulfate. So, can anyone tell me which method is good for precipitating protein from yeast cell lysate? The most important thing is I want to use the precipitated protein of yeast cell lysate for downstream applications like running protein sample on SDS-page, further purification of recombinant protein using spin column of IMAC and western blotting as well. Thank you.
Using Ammonium sulfate will be better as it does not hamper the activity of the protein. The optimization is up to you.
Why is ammonium sulfate better than the other two methods?How about the precipitation of protein using PEG 6000 and 8000?
Cell lysate has always a high amount of protein so, for the purposes described in your question, my advise would be to use a centrifugal concentrator of 3kDa for getting more concentrated protein, presumably, that you would like to achieve by precipitation.
Dear Budak
Acetone and TCA/acetone precipitation is the best methods. Between them, TCA/acetone is more specific and I use it more and more and find good results. Iuse it for 2DE and enzymatic assay.
If you need these protocoles, please dont hesitate to contact me.
Cheers
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