Purification of a cell membrane protein - (Jul/10/2002 )
The protein I am working with binds to the cell (E. coli BL21(DE3)pLysS) membrane. How should I separate it from the membrane and purify it?
I think you have encountered insoluble protein expression from your E. coli growth. When you protein binds to the bacterial cell membrane you have produced inclusion bodies. Actually inclusion bodies are a good thing since pelleting your bacterial will give you >85% recombinant protein. Try using Sarkosyl it separate your protein from the inclusion bodies. For a more detailed explaination please check the Frangioni & Neel paper from Analytical Biochemistry 210, 179-187 (1993) title of paper: Solubilization and Purification of Enzymatically Active Glutathione S-Transferase (pGEX) Fusion Proteins.
Hope that helps!