receptor binding

Caroline Szymeczek-Seay css at med.unc.edu
Tue Jun 29 12:44:05 EST 1999


Hi,
I'm using whole cells in a receptor binding assay.  I'm a bit worried
about how long my cells are sitting in the incubation buffer prior to my
adding the hot ligand.  (I harvest the cells (3 or 4 different
treatments), wash them 3 times, and then count them.  Depending on the
count, i often have to recalculate my expt, and sometimes the cells are
hanging out for 45 minutes or so during this process.)

So my question is, how long can freshly harvested and washed cells hang
out in incubation buffer (basically PBS plus sucrose) before use?  And
what about freezing a whole cell pellet after harvesting and washing?
Can you do that and still do a whole cell assay the next day?

i know that i could isolate membranes instead, but i'm trying to cut a
step here.  i'm not having any problem with NS binding, so i'm happy
with whole cells if i can make this work.

thanks in advance,
caroline




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