I have ran an elisa assay with one side coated with anti igg and one side coated with SEA protein,.,.i put standads in both sides,.but only the side coated with anti igg showed to have working standards but not the site coated with SEA. secondary antibody is the same anti igg conjugate ap,.
Hi Godfrey, I'm afraid that I do not fully understand the description of your assay.
My understanding is that you have an antibody that binds to your protein of interest (SEA protein).
You coated your SEA protein and anti-IgG as a control but only the anti-IgG lanes were positive, is this right?
A common problem with doing direct binding ELISA is that a monoclonal Ab against a protein of interest might not recognize plastic bound proteins because the epitope is hidden when the protein binds to plastic.
I have figured it out.,.the Igg standards will only bind to anti igg..not sea or any protein I use..therefore. If I wanted to coat a plate with a different protein. I will have to get a standard for that coating antibody
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