Problem with ELISAs

Michael Meredith michael.meredith at conted.ox.ac.uk
Mon Nov 16 10:31:13 EST 1998



mark wrote:

> Robin Blumenthal wrote:
> >
> > We are having an interesting problem in our lab with our ELISAs. I was
> > wondering if anyone out there might have any ideas.
> >
> > We are doing sandwich ELISAs for cytokines and Igs (both human and murine)
> > using a biotin-linked secondary Ab, strep-HRP and OPD as a substrate.  The
> > problem we are having is that the developing has slowed down incredibly.
> > Assays that once took 5 min to develop are now taking over 30 min and
> > still not coming all the way up.  We are in the process of testing all the
> > components but in a latest test two different batches of HRP-strep gave
> > the same results.  We are a bit stumped.....anyone have any ideas?
> >
> > We would appreciate any ideas!
> >
> > Thank you
> >
> > --
> >
> > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
> > Robin Blumenthal, Ph.D.
> > Department of Pediatrics, Rm S303
> > Stanford University
> > 300 Pasteur Drive
> > Stanford, California 94305
> >
> > robinb at leland.stanford.edu
> > (650) 723-8224    (650) 498-4632 FAX
> > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
>
> My first guess is PH

  I agree check all of you buffers, we had the same problem and this cured it!




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