[Histonet] mouse on mouse immuno help!!!

[Elizabeth Chlipala]

Amy

If we are performing BrdU on mouse tissue we use a primary from Accurate
Chemical and Scientific (800) 645-6264, Clone BUI/75 product number MAS
250.  It's a rat anti-BrdU.  Then we use a secondary from Dako - rabbit
anti-rat (mouse absorbed) Catalog Number:  E 0468.  I will only use
mouse on mouse antibodies when absolutely necessary, and if I have to do
that I'll use either the Dako ARK kit or the Vector MOM Kit.  Both will
work fine.  I can't send attachments to the histonet so I'll e-mail you
separately with our protocol.

Hope this helps

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)
Premier Histology Laboratory, LLC
P.O. Box 18592 
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
lizchlipala <@t> premierhistology.com
www.premierhistology.com
 
Ship to Address:
Premier Histology Laboratory
University of Colorado
MCBD, Room A3B40
Boulder, Colorado 80309
 
_________________________________ 
This email and any files transmitted with it may contain PRIVILEGED or
CONFIDENTIAL information and may be read or used only by the intended
recipient. If you are not the intended recipient of the email or any of
its attachments, please be advised that you have received this email in
error and that any use, dissemination, distribution, forwarding,
printing, or copying of this email or any attached files is strictly
prohibited. If you have received this email in error, please immediately
purge it and all attachments and notify the sender by reply email or
contact the sender at the number listed above if one is provided. 
 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amy
Porter
Sent: Tuesday, March 16, 2004 2:52 PM
To: Histonet
Subject: [Histonet] mouse on mouse immuno help!!!

Hopefully someone out there can give me some assistance.  I am trying to
stain mouse tissue with a Monoclonal anti - BRDU. We are using a
biotinylated rabbit anti-mouse F(ab')2 secondary at 1:500 in a
commercial diluent with normal rabbit serum added to it.  Our negative
system control (no primary) is coming out almost perfectly clean -
however the slides with primary applied are a whole different story.  I
am seeing what i believe to be good positive signal - with lots of
noise.  Does anyone have any suggestions that might not be working with
commercial kits that are out there.  My lab is research and we don't
usually work with kits.  Just questioning things like - should I
avidin/biotin block all mouse tissue?, should i use low or high
percentages of normal serum in my primary and secondary?.....  I am so
close to having this thing worked out any help would be appreciated.


Amy S.Porter, HT(ASCP) 
Michigan State University 
Department of Physiology 
Division of Human Pathology 
College of Human Medicine 
portera203 <@t> yahoo.com


Do you Yahoo!?
Yahoo! Mail - More reliable, more storage, less spam
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet