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Re: [ccp4bb] Removal of bacterial Hsp70 contaminant from recombinant protein
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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999
Previous message:
Subject: Re: Differences in R and Rfree
From: HKim {- at -} DECODE {- dot -} COM HKim {- at -} DECODE {- dot -} COM
Date: 2007-03-09		Next message:
Subject: Re: Differences in R and Rfree
From: "Santarsiero, Bernard D {- dot -} " bds {- at -} UIC {- dot -} EDU
Date: 2007-03-09


Subject: Re: Removal of bacterial Hsp70 contaminant from recombinant protein
From: Dr shekhar shekhar {- at -} CDFD {- dot -} ORG {- dot -} IN
Date: 2007-03-09

Hsp70 is very hydrophobic, and the problem mentioned by you is quite
common in many affinity based purifications. I suggest that you load your
sample on an HIC (say, phenyl sepharose) and elute with a gradient from
high-salt to no-salt. Hsp70 typically elutes close to no-salt buffers on
the HIC chromatography.

Shekhar Mande

On Fri, 9 Mar 2007, Yong Tang wrote:

> RE: Removal of bacterial chaperone Hsp70 contaminant from recombinant
> protein preparation
>
> Dear all,
>
> I have a protein expressed at 37C for 3 hours in BL21 DE3 and purified
> with sub-stoichiometric amount of apparent Hsp70 contaminant even
> after exhaustive affinity (GST-fusion or His-tagged), ion-exchange and
> sizing column steps. I would like to know if you have a
> well-established protocol for getting rid of such a contaminant. I
> asked around and was told to try adding ATP at certain(?) stage of the
> purification.
>
> I would much appreciate your input.
>
> Many thanks! –yong @ the Wistar Institute
>
>

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999
Previous message:
Subject: Re: Differences in R and Rfree
From: HKim {- at -} DECODE {- dot -} COM HKim {- at -} DECODE {- dot -} COM
Date: 2007-03-09		Next message:
Subject: Re: Differences in R and Rfree
From: "Santarsiero, Bernard D {- dot -} " bds {- at -} UIC {- dot -} EDU
Date: 2007-03-09
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