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 <CCP4BB@JISCMAIL.AC.UK> On Behalf Of amala mathimaran Sent: Saturday, 28 December 2019 12:30 AM To: CCP4BB@JISCMAIL.AC.UK 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
