You want to drain the blocking reagent so that it does not dilute your
primary antibody, you don't rinse because you want it to still be there
taking up the non specific binding sites while your primary binds to the
specific sites.
Patsy Ruegg

-----Original Message-----
From: Montina Van Meter [mailto:mvanmete@postbox.acs.ohio-state.edu]
Sent: Tuesday, March 21, 2000 7:28 AM
To: histonet@pathology.swmed.edu
Subject: IHC question


Dear Histonetters:
		Can someone provide information as to why most IHC protocols
have you
drain off the blocking solution before adding the primary and not rinsing
with buffer in between?  In all the procedures I have looked at they
explain the theory behind the other steps but not this one.  Thanks in
advance.

						Tina


Tina Van Meter
Dept. of Neuroscience
The Ohio State University
Columbus, Ohio