Thank you all for the suggestions. I would be setting up a new crystallization trial in a couple of days and will surely incorporate all the advices.
Thanks once again Cheers!! -Nazia On Sat, Nov 23, 2013 at 11:07 PM, Nazia Nasir Phd2009,ProteinCrystall.Lab < nazia.nasi...@nii.ac.in> wrote: > Hello everyone, > > I am trying to crystallize the truncated version of a memebrane protein > (30 amino acids truncated, including signal sequence a transmembrane > helix). The protein expresses in inclusion bodies and I solubilize it in 8 > M urea and finally purify it in 20mM Tris, 50mM- 300mM NaCl, 5% glycerol, > pH 8.5. I have been able to gow crystals but all of them grow as thin > plates. The best resltion obtained so far is 3.7 A. Moreover, for SAD > phasing using the natural Sulpurs, the Cr beam gives a maxmum resoltion of > 7 A. I have exhausted all screens in my lab and set up expansions of all > positive hits, but all I get are plate like crystals with poor resoltion. > > Can anyone suggest how to improve my crystals? Moreover, is the data I am > getting so far workable for SAD phasing? mY protein is 500 amino acids long > with 11 Methionines, spread out almost through out the protein length. > > Eagerly awaiting some help! > > Thanks ! > > > > > -- > Nazia Nasir > PhD Scholar > Protein Crystallography Lab > National Institute of Immunology > New Delhi > -- Nazia Nasir PhD Scholar Protein Crystallography Lab National Institute of Immunology New Delhi
