crude enzyme precipitation is inclusion body? (View forum version)



Biogareth

Posted 16 November 2011 - 12:48 PM

Recently I tried to express one protein from mushroom into E.coli. I tried to extract crude enzyme from pellet by lysozyme, and got the supernatant by centrifugation. But when I kept the supernatant overnight at 4 degree, there are many precipitation. I wonder these precipitation are inclusion bodys?
If it is the case, how to solve that?
Any suggestions are welcome!

Biogareth

mdfenko

Posted 21 November 2011 - 09:17 AM

not likely. the inclusion bodies (if any) would have pelleted when you centrifuged.

the precipitate upon standing is probably denatured protein aggregates.

what are the buffering conditions of the solution?

Biogareth

Posted 21 November 2011 - 12:44 PM

not likely. the inclusion bodies (if any) would have pelleted when you centrifuged.

the precipitate upon standing is probably denatured protein aggregates.

what are the buffering conditions of the solution?



The solution is:
Tris-HCl (50mM pH 7.5), 1mM EDTA, 4mM MgCl2, 50 mM NaCl and 1mg/ml lysozyme