Insoluble TEV Protease - (May/07/2009 )

Hi all,

I'm having some trouble with solubility of TEV protease - I'm expressing a His-Tagged TEV protease in about 1 L of BL21(DE3) E. coli at 30 C using 1 mM IPTG as the inducer for about 4 hours. However, in all samples I've looked at, all the protease is insoluble. I was wondering if anyone had any tricks, outside of lowering the IPTG concentration or the temperature, on solublization.

Thanks a bunch!

Hi

Try first those lowering IPTG and temp....... what ever first then findout . if still ur prtoein is in insoluble then probably ur expression levels are very high or ur Protein it self is a probelm as some of the proteins never form soluble fractions because of their high molecular weight or hydrophobiciy or protein nature.


Regards
Sudhakar
I'm having some trouble with solubility of TEV protease - I'm expressing a His-Tagged TEV protease in about 1 L of BL21(DE3) E. coli at 30 C using 1 mM IPTG as the inducer for about 4 hours. However, in all samples I've looked at, all the protease is insoluble. I was wondering if anyone had any tricks, outside of lowering the IPTG concentration or the temperature, on solublization.

Thanks a bunch!