Protein precipitation - acetone?

Pepa Florez Pérez via methods%40net.bio.net (by snipeurope from hotmail.com)
Mon Mar 15 05:17:42 EST 2010


Hello Stan,

 

I am sorry about your problems using speed-vac. I am used to concentratre samples containing 150mM NaCl, 0.1% NP-40 even 10X and if I boiled them longer (10 minutes) and run the gel slowly at the beginning it works fine.

I don't know, but maybe it's worth to give it a try!

 

Regards,

 

Eva 
 
> Date: Sun, 14 Mar 2010 19:35:25 -0700
> From: stanleycheungkk from yahoo.com
> To: methods from magpie.bio.indiana.edu
> CC: 
> Subject: Re: Protein precipitation - acetone?
> 
> Hi there,
> 
> I would not recommend concentrating samples by lyophilization or speed-vac. Both concentrate salts, buffers and detergents. I tried to concentrate conditioned medium for doing SDS-PAGE. The samples were concentrated about 10 times. However, the ppt was not fully re-solublized. I don't know what happened to the protein, the result was I saw nothing on the membrane. I believe the solubility of proteins and salts concentration are the problems.
> 
> Best,
> Stan
> 
> 
> 
> ________________________________
> From: lautys <lautys from gmail.com>
> To: DK <dk from no.email.thankstospam.net>
> Cc: methods from magpie.bio.indiana.edu
> Sent: Fri, March 12, 2010 9:33:15 AM
> Subject: Re: Protein precipitation - acetone?
> 
> dear all,
> 
> how about lyophilize low protein concentration sample and resuspend in lower
> volume of buffer/water? is that recommended for protein concentration?
> 
> regards,
> Lau
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