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Dear Amro, the very first thing is that you are very close to your pI (7.8) which should preferably be avoided as it will cause the charge neutralization and your protein will become more hydrophobic. My first suggestion will be to skip the pH value in the range of +/-1 from your pI. it will help you. Second suggestion would be to use sucrose and arginine in your buffer. Your salt concentration sounds to be fine. You can screen for different ranges of buffers. Best -YSR ####@@@@#### Yogendra Singh Rathore On Wed, Jul 4, 2012 at 4:17 PM, Anuradha Balasubramanian <[log in to unmask]>wrote: > check this article its quite useful > > http://www.ncbi.nlm.nih.gov/pubmed/12711344 > > > On Wed, Jul 4, 2012 at 4:09 PM, amro selem <[log in to unmask]>wrote: > >> hallo every body, i face problem with protein when i make gel filtration >> or concentrate my protein . i notice sever aggregation . i use only >> glycerol as additives to 50 mM TRIS and 150 mM NaCL ; PH 8 . my IP is 7.8 . >> any suggestions. >> best regards >> >> Amr >> >> >> >> >> >> >> > > > -- > B. Anuradha > Research scholar, > CAS in crystallography and Biophysics, > University of Madras. >