His-tagged protein, urea & antibody production

Frederik Boernke boernke at ipk-gatersleben.de
Fri Jan 30 14:00:35 EST 1998


Glenda Lawrence wrote:
> 
> Hi.
> 
> Does anyone have ideas or experience re- the best
> way to remove urea before injecting recombinant
> protein into rabbits for antibody production?
> How much urea can be tolerated?
> The protein is a His-tagged protein (insoluble) purified
> and eluted in 8M urea, 20mM Tris, 150mM NaCl and 200mM imidazole.
> 
> Thanks,
> 
> Glenda Lawrence

Hi Glenda,

we usually run a preparative SDS-PAGE (3 mm thick)and cut out the
respective band
after reversible staining with KCl. The protein is electroeluted from
the acryl 
amide using some BioRad apparatus. This works very well and yields
sufficient pure
protein for immunisation.

Ricky

******************************************************************

Frederik Boernke
Research Group of  Molecular Plant Physiologie
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
e-mail: boernke at ipk-gatersleben.de
http://www.ipk-gatersleben.de



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