"Thorsten Schmidt" <Thorsten.Schmidt at rz.ruhr-uni-bochum.de> wrote:
>> Dear Reader!
>> I have big problems with ELISA.
> The main problem is that the color does not develop satisfactorally.
>> I use as second antibody horse-radish peroxidase (HRP) coupled
> anti-rabbit antibodies from donkey (Amersham).
> To develop the color I use a solution of OPD.
> (ortho-phenylenediamin-dihydrochloride)
> in 10 ml 0,05 M Citrate pH 5,2:
> + 40 mg OPD
> + 15 µl 30 % H2O2
>> Some of my experiments might have shown that H2O2 might be an important
> factor for color development.
>[snip]
>> It would be too kind of you if you´ll send me an answer!
>> Thank you in advance
>> Thorsten Schmidt
>You mention H2O2 as an important variable so I assume you
are already using fresh reagent here. Another component that
can 'go off' is the citrate - even as solid. You may want to
try a freshly opened container.
Keith Leppard
