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
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crude enzyme precipitation is inclusion body?
Started by Biogareth, Nov 16 2011 12:48 PM
inclusion body
2 replies to this topic
#2
mdfenko
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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?
the precipitate upon standing is probably denatured protein aggregates.
what are the buffering conditions of the solution?
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#3
Biogareth
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Posted 21 November 2011 - 12:44 PM
mdfenko, on 21 November 2011 - 09:17 AM, said:
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 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
