Can anyone help me about how to improve the sensitivity of the Elisa assay I'm trying to optimize?
I'm coating directly my mAb, saturate, then add samples or standard, after a polyclonal antibody and at last antibody labelled with HRP.
(with washing at every steps)
Do you think I need to increase the incubation time of one of the reagents? or increase the quantity of reagents?
I don't really know what to change to have a better sensitivity.
And another question is about the purity of the protein used as the standard in my test. Do you think a raw recombinant protein is enough or it is better to have a purified protein?
Thanks for your help