发信人: bubba (努比), 信区: Biology
标 题: Re: TCA precipitation
发信站: The unknown SPACE (Wed Mar 27 16:49:48 2002) WWW-POST

hehe, never boil that yellow sample, or you risk degrading your sample.
the yellow color comes from bromophenol blue which turn yellow under acidic pH
and protein will be degraded under acidic condition.
Put extra tris base in your SDS-PAGE buffer to neutralize residual acid. Don't
worry about the undissolved pellet. TCA-precipitated protein should be easily
dissolved by boiling in SDS buffer. Undissolved stuff is likely to be other
stuff.
【 在 bowl (一碗稀饭) 的大作中提到: 】
: I was doing a TCA protein precipitation. After spinning down the pellet, I
washed it with ice-cold acetone. I put SDS-PAGE buffer into it before the
pellet was completely dried. The page buffer turned into yellow instantly and
the pellet doesn't
: dissovle. Can any one help? Or give me a detailed protocol.
:
: Thanks.
:
:


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