his-tagged protein purification contamination

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Mon Sep 25 11:46:24 EST 2000

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

Have you tried to run an imidazol gradient to get a higher purity of the
desired protein?
Maybe your contamination elutes earlier......

Just an idea....
Regards, Ina

Daun Clizbe wrote:

> Greetings
>         We are currently using Clontechs Talon resin (Co++ based) to
> purify a his-tagged protein from cell extracts. The tagged protein is
> eluted with a final concentration of imidazole at .5 M. In using the
> purified protein in downstream reaactions we have found that we
> co-purify either a nuclease or a chemical component that catalyzes RNA
> degradation. Has anyone had similar problems with this product even with
> multiple stringent washes?  Has anyone ever tried DEPC treatment of the
> resin (the company is uncertain of the effects......)?
> Thanks

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

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