Hello
I have been trying to get C-terminal SH2 domain of the p85 protein
for my experiment. Here, I cloned the protein into pet42a as a GST
fusion protein. It is soluble expression and sticking to GST coloumn
very well, and I can purify the protein in huge amount. I eluted the
protein from the coloumn then get ready for the cleavage of GST from
the C-SH2. When time comes to cleave the C-SH2 protein from GST, it
starts becoming problem. Cleaved C-SH2 becomes precipitated in the
solution after cleavage, however cleaved GST stays soluble. I do not
how to get this CSH2 in its soluble form. If you have any idea, I
would be pleased to hear as soon as possible.
Sincerely
Mehmet Sen