coupling proteins to CNBr activated sepharose...

Sai siyer at bmg.bhs.uab.edu
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 bmg.bhs.uab.edu
graduate student, Dept of Biochemistry and Molecular Genetics
University of Alabama at Birmingham...



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