Pharmingen IL-4 Sandwich ELISA - High Background Probs

[J Matthew Buchanan]

I am using the Pharmingen sandwich ELISA (11b11 capture, BVD6-24G2
biotinylated detecting) and am having problems with real high
background.  I have followed a copuple different protocols with
different blocking agents, washes, etc. (including Pharmingen's
protocol word for word), and still have the problem.  I am getting
O.D.'s of about 0.6 - 0.7 for background at 10-20 minutes after
adding substrate.  I have dealt with Pharmingen's tech support
and this has only added problems.  They gave me a different lot #
of both coating and detecting Abs, but the problem remains.

Is anyone else using this system, and if so, are you having similar
problems?  Any solutions?  The plates we are using (COSTAR) work
fine for our other cytokine ELISA's.

Any advice is appreciated.


MAtt


jbuchana at magnus.acs.ohio-state.edu
MBUCHANAN at opus.mco.edu                  - personal
METZGERLAB at vortex.mco.edu               - lab

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