I want to analyze certain cytokines in plasma samples and coat ELISA plates with monoclonal antibodies specific to the cytokines, how do I go about preparing my ELISA microplate ie. from coating the microplates for each cytokine followed by various washing steps. I am looking to employ the quantitative sandwich ELISA technique. How do I validate my protocol and verify my results?

Any insights on the above are highly appreciated.

Yes, I could buy separate human cytokine ELISA kits but my sample size is over 150 and buying separate Cytokine ELISA kits would be super expensive.