coupling proteins to CNBr activated sepharose...

Sai siyer at
Tue Nov 19 15:20:22 EST 1996

hey all,
        anybody here try to couple really diluted protein samples to the
CNBr activated sepharose 4B resin ?  what i mean is that the protocol by
pharmacia recommneds that the coupling solution be really concentrated
(about 5 mg or protein in a 1-2 ml coupling reaction) and this may not be
too feasible for some of us (i.e me) who have proteins that dont like
being too crowded (aggregration in hi concs.).  so i was thinking to
dilute out my coupling reaction with 10 mg or protein in a 30 - 40 ml
solution.  that way, i dont have to worry about aggregation problems
during the coupling reaction.  so would dilution to that much of an extent
affect coupling efficiency??  any ideas?  thanks...

Sai Iyer
siyer at
graduate student, Dept of Biochemistry and Molecular Genetics
University of Alabama at Birmingham...

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