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Dot-blot ELISA - (Mar/30/2008 )

I am trying to standardize a dot bot ELISA using polyclonal Ab, which i raised. I have to do this work for my employer and supply it to him, so they can use the test to detect their Ag efficiently with less time. I did the experiment several times but..unfortunately i end up with very high background. Recently i have found Mab for the same and it doesnt give any background and works fine. how can i make my pAb more efficient compare to MAb. or i should just forget the PAb and start using Mab, develop the ELISA and supply it to employer.
pls help!!


please check the dilution of polyclonal antibody you are using
may be you are using too high concentration because it is known that for very high concentration of polyclonal antibody non-specific binding is supposed to happen
all the best