Purification od insoluble protein (TIMP3) - (Dec/20/2010 )
I attempted to purify TIMP-3 protein, using protein-A sepharose and elutiing the purified protein with sodium citrate buffer pH3. Then, nutrilizing of eluted protein with either Glycine-OH or Tris-HCl buffer pH9. the protein that eluted and nutrilzed by this way provides cell toxixity. So, Could anyone help me in finding alternative buffers that be safe for cells?
Hola , if the furification method is good, I would dyalize against PBS, filter by 0.22um and would add to the cells. Buena suerte y Feliz Navidad
This protein is highly basic, so it should be eluted with low pH buffer and consequently neutralized at high pH, otherwise TIMP-3 will stick into the tube. I tried to concentrate the protein but I lost most of the protein. Could you guide me to find safe alternatives to the elution and neutralizing buffers.