[Histonet] mouse on mouse immuno help!!!
Amy Porter
portera203 <@t> yahoo.com
Tue Mar 16 15:51:51 CST 2004
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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