Ann -

For whatever it's worth, here are my thoughts on your problem.
Do you use Methanol in your peroxidase block (if you use one)?  If so, that
is the culprit.  Change to 0.03%  H2O2 in PBS.  Also, I am assuming since
you are doing antigen retrieval that you are using formalin fixed, paraffin
processed tissue.  Most CD4 antibodies don't work in formalin fixed tissue.
Check with the manufacturer.







"ANN MARUSKA" <amarusk1@FAIRVIEW.ORG> on 08/11/99 09:14:20 AM

To:   histonet@pathology.swmed.edu
cc:    (bcc: Margaret Gondo/GeneMedicine)
Subject:  CD4




Help!
I have been trying to work up CD4, Ventana predilute(Novacastra) clone 1F6
on the NexES with NO success.  I have tried a steam retrieval for varying
times with EDTA bufffer (ph8.0) using tonsil fixed in both NBF and B5.  I
have tried both the basic DAB and Enhanced kits with and without
amplification.  I even thought perhaps my first antibody was not good and
have tried it with a second predilute.
Any help or suggestions would be most appreciated!
Ann Maruska
amarusk1@fairview.org
Fairview-University Med Ctr
Mpls, MN
612-672-4005