Protein precipitation - acetone?
(by shifalich from rediffmail.com)
Sun Mar 14 06:15:48 EST 2010
How about small peptides and proteins? I heard Acetone pptn. loses them.On Fri, 12 Mar 2010 05:53:59 +0530 wrote>In article , "Iraz Toprak Aydin" wrote:>>>>I have to do a western blot, but my protein concentration is very low, and I>>have to run a mini gel. So I was thinking of precipitation the proteins. I>>have never done this before. Does acetone have a bad effect on the blotting?>>Are there any points that I should be careful about?>>Aceton is not very good at precipitating very dilute proteins. Also, >recoveries also vary a lot between proteins. Without a question, >the way to go for you is chloroform/methanol precipitation. >>Ref.: Wessel, D. and Flu"gge, U.I. (1984) Anal. Biochem. 138:141-143.>>Work very well. Only change I'd suggest is to spin HARD (like >10-15 min at 16,000-20,000 g at a step of collecting protein on >the interphase. >>DK>_______________________________________________>Methods mailing list>Methods from net.bio.net>http://www.bio.net/biomail/listinfo/methods>
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