[Histonet] RE: Ki-67 MIB-5 on FFPE mouse tissue

[C.M. van der Loos]

Dear Julie and Liz,
How about using the anti-Ki67 rabbit monoclonal (clone SP6, 
LabVision/Neomarkers)? This antibody worked in my hands on human, mouse 
and rat tissue very well! And you don't have this mouse-on-mouse 
problem. Just 15 min HIER in EDTA9.0 and a polymer/HRP detection.
Hope this helps.

Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands

phone:  +31 20 5665631
fax:    +31 20 6960389
e-mail: c.m.vanderloos <@t> amc.uva.nl 

----- Original Message ----- 
>From  Elizabeth Chlipala <liz <@t> premierlab.com> 
Date  Thu, 13 Jan 2005 15:22:32 -0700 
To  "'Randolph-Habecker, Julie'" <jhabecke <@t> seattlecca.org>, 
histonet <@t> lists.utsouthwestern.edu 
Subject  RE: [Histonet] Ki-67 MIB-5 on FFPE mouse tissue 

Julie

The Ki-67 from Dako that works on mouse tissue is the rat anti-mouse
Ki-67 (clone TEC-3), cat # M7249. The MIB-5 works on rat tissue.  As far
as I am aware these Ki-67 antibodies from dako do not cross react with
other species.  For Ki-67 I use three different antibodies, one for
human, one for mouse and one for rat.  I have tried the human on rat and
mouse and I know that does not work.  I'm assuming the MIB-5 will not
work on mouse, plus it's a mouse monoclonal.  I normally try to stay
away from mouse on mouse if other antibodies are available.  I use the
rat anti-mouse Ki-67 at 1:50 with steam retrieval.  I have images and a
more detailed protocol if you would like

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com
 
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Randolph-Habecker, Julie
Sent: Thursday, January 13, 2005 2:49 PM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] Ki-67 MIB-5 on FFPE mouse tissue

I have another question.

I am trying to get Dako's Ki-67 MIB-5 working on FFPE mouse tissue. I
have
heard that it works on mouse tissue and it works great on FFPE human
tonsil.
Does anyone have a great protocol for mouse?

Thanks!

Julie

Julie Randolph-Habecker, Ph.D.
Experimental Histopathology Shared Resources
Fred Hutchinson Cancer Research Center
1100 Fairview Ave. N, G1-300
PO Box 19023
Seattle, WA 98109-1024
Tel: (206) 288-1187
FAX: (206) 288-1345
jhabecke <@t> fhcrc.org <mailto:jhabecke <@t> fhcrc.org>