The cell lysate shows my protein but, after nickel nta it degrades and i get a 54Kda band.But, the size of my protein is 64Kda. Maybe the nterminal is surviving while the c-ter disappears.
I have used pmsf before lysis.
wash and elution have 500mM NaCl. It is a eukaryotic protein, i use rosetta gammi 2 for expression at 1.5mM IPTG(22C) induction.
Adam B Shapiro
Consider using a protease inhibitor cocktail instead of relying only on PMSF.
2 votes
1 thanks
Stefan Gajewski
64k Band im your lysate could be an e. coli protein. Can you confirm your protein runs at expected size on SDS? E.g. western blot?
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Proteins
More Subhash Narasimhan's questions See All
Similar questions and discussions