MS on gel-isolated proteins

johnp at GGPL.ARSUSDA.GOV johnp at GGPL.ARSUSDA.GOV
Mon Apr 8 11:57:12 EST 1996


>Date: 3-Apr-96 12:26:
>  From: treefrog at u.washington.edu (Bryce Chackerian)
>
>Does anyone out there have a good protocol for preparing a glycoprotein
>for Mass Spec. by excising a band from an acrylamide gel?  Apparently SDS
>is not good when doing MS.
>pls e-mail treefrog at u.washington.edu
>Thanks in advance
>-bryce
>


Bryce:

Pierce Chemical Co. sells a product called "Extracti-Gel D Detergent Removing 
Gel (Product #20346 for pre-packed columns) which they say will remove SDS 
(and other detergents) by an affinity purification without low non-specific 
binding of the protein.  I have not used this product, and I have no 
connection with the company, but you might call them and ask them about its 
application to your use.  Their technical service people are generally very 
good.  Their telephone number is 800-874-3723.  You might also be able to 
separate your protein from SDS by electroelution through an appropriately 
sized dialysis membrane (one that would retain your protein, but let the SDS 
migrate to the anode).  This would have the added advantage of eluting the 
protein from the gel slice and separating it from the SDS in one step.  
Again, I have not done this myself.  Someone else may be able to comment on 
whether these approaches work in practice.

Good luck.

John



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