Quick navigation:        Home   |    Site Map   ||    References   |    Biography   ||    Copyright   |    Other copyright   |    Contact us   |   
Protein structure
 

[ccp4bb] S-tagged protein purification
 

Basic tutorials: 		 
 

CCP4bb navigation

CCP4bb <-- 2007 <-- May 2007 <-- 12 May 2007
Previous message:
Subject: Low resolution model building
From: "Kolstoe S {- dot -} E {- dot -} " S {- dot -} E {- dot -} KOLSTOE {- at -} SOTON {- dot -} AC {- dot -} UK
Date: 2007-05-12 		Next message:
Subject: Re: Superdex 200 PC columns
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-05-12


Subject: S-tagged protein purification
From: "Y {- dot -} -F {- dot -} Li" yongfuli {- at -} GMAIL {- dot -} COM
Date: 2007-05-12
Hi,

I have a membrane protein with a C-terminal S-tag, and there is no cleavage
site for the S-tag (novagen, e.g, thrombin, EK). I wonder whether I still
can use this for purification without cleavage of the S-tag. Here are my
questions:

1) How to release the protein bound to S-tag resin? I noticed they suggest
3M NaSCN, 3M MgCl2 or 0.5 M Citrate pH2, but these sound crazy for a
membrane protein. Does anyone have experience with this?

2) It seems that S-tag resin has lower binding capacity than does Ni resin.
Did anyone ever use S-tag resin for purification of at least 10 mg protein?

I would probably put a his tag on my protein for purification later.

Many thanks for any suggestions.

Yong

CCP4bb navigation

CCP4bb <-- 2007 <-- May 2007 <-- 12 May 2007
Previous message:
Subject: Low resolution model building
From: "Kolstoe S {- dot -} E {- dot -} " S {- dot -} E {- dot -} KOLSTOE {- at -} SOTON {- dot -} AC {- dot -} UK
Date: 2007-05-12 		Next message:
Subject: Re: Superdex 200 PC columns
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-05-12


ProteinCrystallography.org: Copyright 2006-2007 by Quid United Ltd