Solubilization of a His-tagged protein.
yoel and adaia shiboleth
shibolet at mishkei.org.il
Sat May 27 02:44:22 EST 2000
try growing the cells at 20 C and lower concentrations of IPTG (0.6mM).
worked for me!
Fernando Touris Otero wrote:
> Hi all!
> I´m trying to solubilize a His-tagged fusion protein. I did solubilize
> it once although I wasn´t able to reproduce it. What I did was to grow
> the bacterial cells until they reach an OD= 1.0 and after an induction
> period of 3 hours growing at 30C (1 mM IPTG), I followed a typical
> solubilization protocol for inclusion bodies, including a washing step
> in Triton X-100 and several incubations from 2 to 8 M urea.
> I already tried using 6 M Gd-HCl. (and growing the cells at 37C during
> induction it doesn´t help).
> Does anyone know anything to help me out?
> Thanks a lot :)
> Celina Costas.
> Universidad de Santiago de Compostela (Spain).
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