Isolation of a receptor using mAB
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.
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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