[Histonet] mouse on mouse immuno help!!!

[Gayle Callis]

AMy, 

YOu are doing mouse on mouse immunostaining and you should use a kit
designed for that use.  Our research lab normally does not work with kits
until it comes to frustrating mouse on mouse staining. That is when we go
commercial. The DAKO ARK kit should work, Chemicon has a new kit that works
well, there are others on the market.  

The kicker with these kits is the special blockers they supply in order to
reduce the mouse to mouse detection, aka high background.  Another kit that
we have used is Scytek out of Logan Utah.  I was surprised (I am a terrible
doubtful type when it comes to ms on ms staining!) at how clean the new
Chemicon kit was, simple and easy to use. 

At 01:51 PM 3/16/2004 -0800, you wrote:
>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
>
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)