Hello everybody,
I just purified a membrane protein tagged with GFP, which has a cleavage site
of preccission protease. And I got a problem with removing the GFP tag by
preccission protease (1mM DTT and 1 mM EDTA were included in the buffer). It
can not cut my protein. I have already tried to digest it in different
detergent like DDM and C12E8 (also different concentration for detergent like
lowering the detergent to 1.1 x cmc since a recent science paper lowered the
detergent to this level and got it worked). I know this depends on the
different proteins. I am wondering if anyone has this experience in digesting
membrane protein by preccission protease. Any suggestions are appreciated.
Otherwise I might have to go back to just his-tag if there is no trick for
that. Thank you so much in advance.
All the best
Qiangmin Zhang
