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Recommended maximum OD values in ELISA optimization. What's the reason?

Recommended maximum OD values in ELISA optimization. What's the reason? - ELISA Assay Forum

Recommended maximum OD values in ELISA optimization. What's the reason? - ELISA Forum. Discuss and post questions including optimization, troubleshooting, protocols and theory of ELISA assays using monoclonal, and polyclonal antibodies or peptides.


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Old 04-23-2012, 10:43 PM
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Question Recommended maximum OD values in ELISA optimization. What's the reason?



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!
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competitive elisa , elisa , maximum , od values , optimization , reason , recommended , values


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