No endpoint in Indirect Peptide ELISA! - (Apr/10/2005 )

Bioforum,
I am new to this forum. Please help if possible... I am running an indirect peptide ELISA with very concentrated antibody taken from the serum of a hyperimmunized pig. I have found that gradations in signal occur when I am making the primary Ab dilutions up to 1/50,000. However, from 1/50,000 on to 1/10,000,000, I have a plateau effect where the signal remains the same. I am unable to achieve a signal similar to the negative controls. The signal remains two times as high as the negative controls. Any ideas?? Thanks for your time, hood

Answering my question...

I am examining the possibility that my polyclonal goat anti-pig peroxidase labeled antibody (KPL) is recognizing my synthetic viral peptide giving me a constant, albeit low signal irrespective of the dilution of my primary antibody. I'll have the data tomorrow...