Dear all.. Wow so many answers.. really thanks to all I will fellow your ways one by one..
Thanks and regards Amala On Sat, Dec 28, 2019 at 9:31 AM Newman, Janet (Manufacturing, Parkville) <[email protected]> wrote: > Hi, > > > > To add to this thread, there are a few more easy things to try – > > > > Try doing matrix microseeding and try doing limited proteolysis. Even > though the following link describes how we do this in our laboratory, (the > C3 Facility in Melbourne) the pages give a quick overview to both matrix > microseeding and proteolysis, and provides links to a recipe for making > seeds etc. > > > > https://research.csiro.au/crystal/user-guide/soluble-proteins/ > > > > Then you might want to try setting up crystallisation at different > temperatures, and using DSF or some other technique to find a different > formulation for your protein, as that can significantly alter the behaviour > of your protein in crystallisation trials. > > > > Cheers, and may your new year’s resolution be better than 2A > > > > Janet > > > > *From:* CCP4 bulletin board <[email protected]> *On Behalf Of *amala > mathimaran > *Sent:* Saturday, 28 December 2019 12:30 AM > *To:* [email protected] > *Subject:* [ccp4bb] how to get protein crystal > > > > Dear all, > > > > Can you suggest me how to get protein crystal??? > > > > I purified my target protein and concentrated to 12mg/ml (pI 6.04). Final > purified protein contains 50mM HEPES pH 8.0 and 50mM NaCl. Then the initial > screening was done using hanging drop method but no crystal. So 2mM NADP > cofactor was added again screen with Hampton (PEG ion, PEG RX, crystal > screen, Index) and Molecular dimensions conditions etc. I got precipitate > like formation the image was attached below. From this formation what I can > do… mean while I increase the protein concentration and did screening for > that selected conditions again I got same kind of formations. I am new to > protein crystallography kindly suggesting me. And how much concentration is > suitable for protein crystallization?? How to find which concentration is > enough for our target protein crystallization?? Thanks in advance > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
