[Histonet] mouse on mouse immuno help!!!

[Amy Porter]

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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