acetone precipitation of samples (for SDS PAGEs)

Hernan Espinoza espinoza at
Mon Nov 18 15:53:57 EST 1996

siyer at (Sai) writes:

>hey all, 
>            i've posted this before, but i figured i'd try again to see if
>there were any other ideas.  i've been doing acetone precipitation to
>remove salts from my samples prior to loading on sds gels.  the basic
>protocol is add 8 vol. of acetone to 1 vol. of protein sample --->

	Different proteins precipitate in different concentrations of
acetone.  I would suggest you try a range of acetone concentrations
(20-80%) and go with the minimum that precipitates your protein.  No
guarantees, but it could help.

	Alternatively, try TCA precipitations.  The wisdom that was
passed on to me was :

	-Add 1/5th volume 100% TCA
	-Incubate on ice 30'
	-Spin 5' in a microfuge (14,000k) @ 4C
	-Resuspend in a minimal volume of SDS Loading Buffer
	 which will turn yellow from the TCA.
	-Neutralize sample with 2M Tris (un - pH'd) so it
	 turns blue again
	-Load that sucker.

	Good Luck. -Hernan

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