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>_______________________________________________>Method!
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