[Histonet] SV40 staining

[Carrie Kyle-Byrne]

Hi Lee,
i'd say that the problem you are experiencing is related to the fixation of
the tissue your collaborator has sent you.  my suggestion would be, and you
may have already done this, to play with the length of time spent in the
antigen retrieval.....perhaps going even longer that what you normally use
for the Cos7 cells (they were fixed overnight in 10% NBF).  Pab101 should
work for you just fine reguardless of species ....it's seeing a virus
protein.....not a protein that is species specific.  (for those not
familiar, Cos7 is a line derived from monkey kidney).
Good luck,
Carrie Kyle-Byrne, BHS, HT(ASCP)
Assoc. Research Scientist II
Antibody Technology Lab
Signal Transduction Research

Exelixis, Inc.
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Email: ckbyrne <@t> exelixis.com

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----- Original Message -----
From: "LD Ridley" <L.D.Ridley <@t> bham.ac.uk>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, October 09, 2003 1:43 AM
Subject: [Histonet] SV40 staining


> Hi all,
>          I'm still relatively new to immunohistochemistry but I seem to
have hit a bit of a problem and would really appreciate any help. My problem
is that I'm trying to stain SV40 T antigen in monkey kidney infected with
SV40 (FFPE)using the Chemate Envision detection kit. I have had success with
staining infected Cos7 cells using the antibody bought from Santa Cruz (Pab
101) sc-147 using extreme antigen retrieval techniques (so i know my
antibody and technique seems fine). However when i try to stain the kidney
slides i cant get any positivity. i've tried everything in vain. The guy
that sent me the block can get nice staining using clone Pab416  My question
really to anyone out there is.. can one antibody work on one tissue but not
on another? and can different clones of an antibody work on one tissue and
not on another even though they are both 'looking for' the same antigen.I
have done a thorough literature search and both clones should detect the
Large T antigen of SV40 so i'm reluctant to give up on Clone Pab 101 before
rushing out to buy clone Pab 416.
> Sorry this quite a long one but any help from anyone working with SV40
would really be appreciated. I'll probably need to clear a few things up
that i havn't explained very well here.
> Thanks in advance
> Lee
> Lee Ridley
> Research Associate
>
> L.D.Ridley <@t> bham.ac.uk
>
> Birmingham Women's Hospital
> Reproductive & Child Health
> Academic Unit 3RD Floor
> Edgbaston
> Birmingham
> United Kingdom
> B15 2TG
>
> Tel: +44 (0)121 6074754
>
>
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