Why is so much of my protein coming out in wash fraction?

I am working on a membrane protein of size 70 kDa. It has a 6X-His Tag on C terminal. I am using 20mM Tris, 200 mM KCl, 10% glycerol, 5 mM beta-mercaptoethanol and 0.03% DDM for the protein...

11 November 2019 1,319 3 View