Thrombin and high salt

nospam at our.site nospam at our.site
Tue Mar 14 13:08:47 EST 2000


This message has been posted by:  612 right <i.hinners at REMOVE-THIS-TO-SENDicrf.icnet.uk>

Hello,
why not cleave on the column?
After loading and washin, you equilibrate against a suitable buffer and
cleave on the column.
saves you another loading step to get rid of the GST.

Hope that helps, Ina


Cyril Privenzentzev wrote:

> Dear Netters:
> I'd like to ask does high salt (up to 1M of NaCl) affect thrombin
> protease activity? I need to cleave GST-fusion protein eluted in 0.9M
> NaCl, 25 mM HEPES pH=7.6, 1 mM EDTA, 0.5 mM PMSF, 1 mM DTT, 10%
> glycerol. I have to get rid EDTA and PMSF, it's clear, but what about of
> salt and glycerol? May be someone know optimal condition for cleavage?
> Thank you in advance,
>
> Cyril V. Privezentz
> CNRS UMR 8532-IGR
> Villejuif
> FRANCE
>
> TEl. +33 1 42115404
> ---

--
Ina Hinners
ICRF
Secretory Pathways Laboratory
44 Lincolns Inn Fields
WC2A 3PX London
UK
email: I.Hinners at icrf.icnet.uk






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