RE: [Histonet] rat gi

From:"Favara, Cynthia (NIH/NIAID)"


You could biotinylated your primary or you could try a rat adsorbed anti
mouse secondary. I believe that Jackson has one made in horse. Sometimes
this will decrease the sensitivity but that can usually be overcome with

Cynthia Favara
903 South 4th Street
Hamilton, MT 59840

The information in this e-mail and any of its attachments is confidential
and may contain sensitive information. It should not be used by anyone who
is not the original intended recipient. If you have received this e-mail in
error please inform the sender and delete it from your mailbox or any other
storage devices. National Institute of Allergy and Infectious Diseases shall
not accept liability for any statements made that are sender's own and not
expressly made on behalf of the NIAID by one of its representatives

-----Original Message-----
From: Till, Renee [] 
Sent: Friday, March 04, 2005 8:35 AM
Subject: [Histonet] rat gi

Does anyone have any suggestions for reducing background with rat gi? I
have been trying to do a PCNA stain on distal and proximal colon with
varying results. We use M0879 PC10 PCNA from Dako, which is a monoclonal
mouse antibody, along with Biotin Conjugated Affinity Purified Goat
anti-Mouse IgG from Rockland, and Peroxidase Conjugated Streptavidin
from Jackson. I block with CAS block from Zymed and antigen retrieve
with CitraPlus from Biogenex. I have also just tried using the
Vectastain ABC Mouse kit, but that seems to turn out even worse than
when I make my link and label. Could it be something as simple as longer
washes or different antigen retrieval? Is it that important that you let
the slides cool for the 20-30 mintues after retrieval as opposed to
continuing right away or cooling them yourself? Or is gi just a tissue
that is always going to give me background problems. Or could it be the
antibody itself? I will admit I don't know enough about the specifics of
how immunos work to figure out exactly what it is that could be causing
the background staining.  So far I have just tried to get them stained
as best as possible without the background staining interfering with
with my counting ( I have to count crypts cells by the intensity of
staining, which is supposed to relate to the cell phase).




Confidentiality Notice: This e-mail message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information.  Any unauthorized review, use, disclosure or
distribution is prohibited.  If you are not the intended recipient, please
contact the sender by reply e-mail and destroy all copies of the original

Histonet mailing list

Histonet mailing list

<< Previous Message | Next Message >>