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.


More information about the Methods mailing list