ELISA HELP NEEDED Cant Get anything to Work

Barbara via methods%40net.bio.net (by blarrain from aol.com)
Fri Mar 23 10:49:05 EST 2007


I'm trying t odevelop an ELISA Assay to measure ENAC & ATPase in mouse
cochlea lysates. I've been able to do so using NFkB, TNF & VEG kits.
But there are no kits for these ab.

I have tried coating the Nunc maxisporp plates ( at a couple of conc)
with rabbit anit-mouse ENAC or rabbit anti-mouse ATPase then add my
antigen (mouse lysate) followed by ALphatase goat anti rabbit IgG. It
seems that nothing sticks. Given the small amount of lysate &
variability coating wit hthat is not practical.

I also tried biotynating the anti ENAC & ant ATPAse & then incubating
with coated plates. Followed by blocking & then my mouse lysate & the
nthe goat anti rabbit. Every thing is high except fo the ones without
the biotinalyted AB initially added.

Anyone have any thoughts or suggstions.

Thanks
BL



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