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Hi all, I am purifying a basic protein with pI 7.8. I used 50mM tris 8.8, 300 mm NaCl, 5% glycerol, 0.1mM PMSFand 5mM beta ME in the buffer and performed affinity purification. Eluted with 200mM imidazole. While elution I could see slight turbid in eluted protein (I get pure protein, single band on SDS-PAGE). However, when I centrifuged I couldn't see any pellet. After some time(a day) protein got degraded. In other method I tried cation exchange purification of lysate with MES buffer pH 6.5. Here also I saw slight precipitation and later degradation. After this again I tried with MES buffer pH 5.5 and PIPES buffer pH 6.0, here also same problem I faced. Please suggest me a method where I can get stable protein. Thanks and Regards, Manjula R Research Scholar Department of Biophysics National Institute of Mental Health and Neurosciences Bengaluru-29, Karnataka INDIA E-mail: [log in to unmask] Mobile no:+91-9538553356 http://www.nimhans.kar.nic.in/ <https://www.nimhans.kar.nic.in/>