Greetings to all, ye Lore Keepers. Ths is my first post.
I'm a postgraduate student in Argentina.
I'm optimizing an indirect competitive ELISA to quantify a small molecule in plant tissue. Reading Crowther's ELISA Guidebook (awfully edited) i've found that when optimizing a competitive ELISA, the recommended antigen concentration to coat plates with is that which results in a maximum OD (plateau) of 1-1.5 for antibody excess. And an antibody dilution which results in 70% of that maximum OD (since competitive ELISAs are reagent limited).
My question is: why not greater OD plateau values e.g. 2.0 (provided we are having the same background OD)?
I'm using a Tecan Sunrise plate reader.
Any ideas? Thanks in advance!