Solubilization of a His-tagged protein.

Fernando Touris Otero bntouris at usc.es
Fri May 26 12:55:42 EST 2000


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).







More information about the Methods mailing list