I tried many buffers in achieving my protein in soluble fraction, fortunately I am successful at pH1. Moreover, I have not used any buffer for pH1: its just milliq adjusted to pH1 using conc HCL. My question is: will I be able to do further studies on this protein with affecting its solubility? Now, it is in the state of impure fraction. My proteins pI is 5.78. So, which purification method is suggested?
Dear Venkat,
The HCl could serve as an "buffer" you equilibrate Superdex column, thus size-exclusion in this weird situation is the first hit I'm finding. Did it many times with HCl, seems to work for proteins like low pH. Anyway I'm afraid your protein could be slightly aggregated, just make sure before loading on the column you spin it down at highest G possible.
Keep fingers cross - Jan
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