His-tagged protein, urea & antibody production

Koen De Smet k.desmet at nospam.ic.ac.uk
Mon Feb 2 03:47:55 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


I don't know if this is the "best way", but we have succesfully produced 
antisera by dialysing the urea away. In most cases, the recombinant 
His-tagged protein preciptated, but that does not matter. YOu can still 
make a suspension with incomplete Freunds adjuvant


Koen De Smet
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