gelfiltration on superdex 75 pg

Sigrid Van Boxstael svboxsta at vub.ac.be
Thu Sep 28 10:41:05 EST 2000


Hi,

I'm trying to purify my protein.
I bring 10 mg (already 90 % pure) on a Pharmacia superdex 75 pg column
and
I can only recuperate 1.5 mg in the obtained peak.
The buffer I used : Tris 20 mM pH 8.2, 2 mM mercapto-ethanol, 150 mM
NaCl.
When I clean the column after the run there is coming protein of it.
I presume that my protein sticks on the column.

I think that increasing the salt concentrations might  be the solution.
Does anybody know either this might help and if yes until what
concentration
I can increase it?


Thanks,
Sigrid







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