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Hi LiuQing, I had a similar case like this before. After I cut the concentration of protein (50%), then I got image_1. To further test the pH, pH 6.5 and pH 7.5 were used. Under pH 7.5, I got image 2. Eventually, the pH were modified to pH 7.8, I got crystal (image 3) with the diffraction of 2.0 Ang. 1. reducing protein concentration. 2. Explore pH range. 3. test different buffer salts with the same pH. 4. optional: adding ZnCl2 or Cd(NO3)2 or (NH4)2SO4: < 5 mM, However, each case is different. I hope this will be helpful to you. Hope to see your publication soon. Kevin ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1