Hi,
There is no way you can compare NiNTA and GSH affinity supports.
It is a lot easier to regenerate NiNTA.
This has to do with the ligand, not the support.
Kd is 10-100 times lower for GSH.
GSH contains a "highly" reactive SH group if enough thiolate is present
(oxydation and/or nucleophilic attack).
It is recommanded to avoid hi pH and the presence of metal ions.
HTH,
Nadir
--
Pr. Nadir T. Mrabet
Structural & Molecular Biochemistry
INSERM U-954
UHP - Nancy 1, School of Medicine
54505 Vandoeuvre-les-Nancy Cedex
France
Tel : +33 (0)3.83.68.32.73
Fax : +33 (0)3.83.68.32.79
E-mail : [log in to unmask]
Selon "Oganesyan, Vaheh" <[log in to unmask]>:
> On the same note with Mirek: does anyone know of a source other than GE for
> resin for purification of FLAG-ed proteins?
>
> Thanks.
>
> Vaheh
>
> -----Original Message-----
> From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of Mirek
> Cygler
> Sent: Tuesday, November 02, 2010 11:47 AM
> To: [log in to unmask]
> Subject: [ccp4bb] Glutathione sepharose
>
> Hello,
> For various reasons we are frequently expressing proteins with a GST
> tag. The glutathione sepharose beads that we are using for affinity
> purification seem to be difficult to regenerate and we see much lower
> capacity when used the second time. We are following the manufacturer's
> instructions for regeneration but this not very effective process as opposed
> to NiNTA which can regenerate multiple times. Due to this, the purification
> of GST-tagged proteins is quite costly. Do you know of a GSH-sepharose that
> can be successfully regenerated several times? Any comments will be greatly
> appreciated.
>
> Mirek
>
>
>
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