Isolation of a receptor using mAB

Leslie Confer lconfer at moose.erie.net
Thu Feb 1 20:11:06 EST 1996


In article <bidinotto.1.24.310EFA1A at postbox.acs.ohio-state.edu>,
   bidinotto.1 at postbox.acs.ohio-state.edu (Andrew Bidinotto) wrote:
>To anyone who may know,
>
>I am trying to isolate a receptor by using a mAB.  I know I could link the mAB 
>to a gel column and run my receptor mix down, but is there a more efficient 
>way of using the mAB to do this.  I thought about attaching the mAB to wells 
>and then appling my extract in the wells, but I am not sure if that will yield 
>enough of the receptors.  If anyone has any ideas feel free to e-mail me at 
>the above address.  I would appreciate it greatly.
>
>thank you
>
>andy b.

I don't think that coating the MAb to the well would be
very efficient.  How about a batch process using your
MAb linked to NHS activated sepharose (or a hydrazide derivative),
but instead of using a column, use a small amount in a microfuge 
tube and allow it to mix (possibly overnight @ 4C).  Then spin 
down the beads, decant supernant, wash and spin X times, and elute.
The beads would be more efficient than wells, providing
greater yields, and the batch process wouldn't use up as
much antibody as required to prepare a column.






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