I don't have much experience with ELISAs, but I do understand that the calibration curves are sigmoidal and linear regression does not give a good r^2 value. In the past I used to use EIA kits from Cayman and they have templates that customers can download for quantitaiton.

I do not have the software to generate a 4-parameter logistic curve fit

Can anyone tell me if I can obtain a general ELISA quantitation template from somewhere?

Also, what curve fit do you use? I have comtemplated using 2-order polynomial but then I am not sure how to apply the equation in Excel to quantitate my unknowns because I am not sure how to use some of the terms, for example:

**y = 2E-09x^3 - 4E-06X^2 +0.0024x + 0.01 (what is the value of E???)**

Any help will be higly appreciated !