Protein precipitation - acetone?

Stanley Cheung via (by stanleycheungkk from
Sun Mar 14 21:39:16 EST 2010

Hi Niko,

Could you describe a bit more about how to make a
acrylamide gel which can compress the protein band (using mini-gel from
Biorad)? I have problem in losing resolution with large sample volume.


From: Nikola Wenta <Nikola.Wenta from>
To: methods from
Sent: Fri, March 12, 2010 1:27:40 AM
Subject: RE: Protein precipitation - acetone?

Hi Iraz!
I don't have any clue about acetone precipitation of proteins, but generally, precipitation with saturated Ammonium sulfate solution provides a good means to concentrate proteins and to get them into a save state for short-term storage. Unfortunatelly, you would already need the protein solution to be at > 1 mg/ml in order to get it to precipitate. Thus, in your case this method doesn't seem suitable. You could also try to concentrate the protein with Centricons, but you would rather loose protein to the membrane than concentrate your solution as it is already too diluted. Why not loading maximum volume into biggest possible pockets on a gel with maximum thick spacers? Additionally you could use a acrylamide percentage that "compresses" your protein band, giving you a better signals in WB.
Best, Niko

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Von: methods-bounces from im Auftrag von methods-request from
Gesendet: Do 11.03.2010 17:03
An: methods from
Betreff: Methods Digest, Vol 58, Issue 7

Message: 8
Date: Thu, 11 Mar 2010 16:56:05 +0100
From: "Iraz Toprak Aydin" <iraz.aydin from>
Subject: Protein precipitation - acetone?
To: <methods from>
Message-ID: <004401cac133$5b7c0df0$127429d0$@aydin from>
Content-Type: text/plain;    charset="us-ascii"

Dear all,

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?

Thanks in advance...

Iraz Toprak Aydin

EPFL SV ISREC, Station 19

Batiment SV, SV 2540  

CH-1015 Lausanne


Tel: +41 21 693 07 36

e-mail:   <mailto:iraz.aydin from> iraz.aydin from


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