This protein does not have any trans membrane helices, as predicted by TMHMM server (http://www.cbs.dtu.dk/services/TMHMM/). Protein is predicted to be outside the cell by TMHMM. This protein is not there in the soluble fraction, but it is in the cell pellet. I will try purification with the detergents. Hope this works. Thanks a lot for help.
On Tue, May 5, 2009 at 6:08 PM, vinothkumar <vku...@mrc-lmb.cam.ac.uk>wrote: > If it is really a membrane protein I am afraid you have to use detergent > for purification. Since you did not use detergent the protein did not elute > of the column, the first thing to try is to solubilize your protein in > detergent (that is from membranes) and purify in presence of detergent. This > way you don't have to worry about refolding. How many TM helices does your > protein has? > > > Vinothkumar K.R. > Structural Studies Division > MRC-Laboratory of Molecular Biology > Hills Road, Cambridge. > CB2 0QH > > Tel: 44-1223-402405 > > > > > > -- Sanjiv Kumar Lab. No. 411, Functional Genomics Unit, Institute of Genomics and Integrative Biology, New Delhi-110007 India
