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help with antibody

Artur Kania ak990140 at imgen.bcm.tmc.edu
Sat Nov 11 12:43:15 EST 1995

hello everyone,

I am looking for help with Ab staining of embryos. I have generated an
antibody that works great on westerns, however in embryos it gives extremely
weak signals (and almost no background). I suspect that the antigen needs
to be denatured in embryos (just like it is in the western blot).
I have been using a standard fixation protocol:  4% paraformaldehyde in 
phosphate  buffer. 

My question :
Is there a way to modify the fixation to include a detergent or something 
which will denature my antigen ?  Should I use this detergent throughout 
the Ab staining protocol ? Has anyone encontered this problem before (and 
solved it) ?

Any help is greatly appreciated 


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