coupling proteins to CNBr activated sepharose...

[Dr. David Martin]

In article <siyer-1911961421520001 at bmg147.cmc.uab.edu>,
   siyer at bmg.bhs.uab.edu (Sai) wrote:
>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...
>

It should go fine.. just make sure you agitate well.

.d

* David Martin - Atherosclerosis and Thrombosis research group  *
* http://www.uio.no/~damartin/      david.martin at biotek.uio.no  *