How to check standard curves in ELISAs - (Apr/19/2014 )

I am trying to understand how standard curves are developed. We use Polynomial 4 point in a computer program. What I am interested in is how do I check my standards to make sure that they ran correctly. What should a cut off be for CV for each standard? Do I have to worry about my actual standards being vastly different from my expected standard values? Then I use R^2 value to make sure that my data still fits my standard curve.

Thanks!

Leelab

Usually for ELISAs, a 4 parameter logistic curve fit is used. You can purchase standard from a commercial source and see how serial dilutions of it compare with your internal standards in the ELISA.