[ccp4bb] S-tagged protein purification

- Protein crystallography

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- Crystallisation

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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

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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 <-- 1999 <-- November 1999 <-- 30 November 1999

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