RE: Making reluctant antibodies work

From:"Turner, Scott"

I forgot to mention that I have tried dilutions of 1ug/mL, 2ug/mL, 5ug/mL,
10ug/mL, 25ug/mL, 50ug/mL and even on a whim tried 100ug/mL and none of them
have worked.  In theory, an antibody and it's antigen should reach
equilibrium after 30min at room temperature, though it might help to try it
at 4 C.  I have tried incubations overnight at 4 C in the past (with
antibodies other than this one) and find that 30min at RT usually yields
better results.

I should also mention that I can't call the supplier, since these are in
house antibodies and unfortunately I usually know more about how they work
in IHC than anyone in that department.

Scott Turner
DNAX Research

-----Original Message-----
From: Swain, Frances L []
Sent: Tuesday, August 13, 2002 2:26 PM
To: 'Turner, Scott'
Subject: RE: Making reluctant antibodies work

Scott, If you will incubate your antibody overnight at refrigerator temp in
a humidifying chamber you will get optimal labeling if there is to be any
labeling. It should work but I thing that either your dilution is wrong or
the time of incubation. 

Frances L. Swain HT(ASCP) A. A. S.
Special Procedures Technician
University of Arkansas For Medical Sciences
Department of Orthopaedic Surgery
Center for Orthopaedic Research
4301 West Markham Street Mail Slot 644
Little Rock AR 72205
Phone (501) 686-8739
FAX (501) 686 8987

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