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Problem with protein precipitation


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#1 iricinforce

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Posted 17 March 2009 - 10:29 AM

Hi everybody,
I have a big problem : I am trying to precipitate proteins from nuclear and cytoplasmic fractions using acetone.
I have unfortunately a problem since I have got a loss of 80 % of my proteins after precipitation. Moreover, the pellet of protein is grey whereas it should be white... Do you know what could be the problem ?
I use a buffer with HEPES 10 mM, MgCl2 1.5 mM, KCl 10 mM, DTT, protease and phosphatase inhibitors and 0.05 % NP40... I don't know what to do...
Thanks a lot for your advices.

#2 lotus

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Posted 19 March 2009 - 01:19 PM

maybe you should try precipitating with ammonium sulphate................




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