How to read ELISA Results?

[joey20]

Hi, how do you ELISA experts read elisa assay results?

i am confused from the data as its my first time. thank you all

[danfive]

You read absorbance on a spectrophotometer.
Calculate average absorbance for each std and sample. Use the avg absorbance for calculations.
Plot Absorbance values of standards only (x-axis) vs Protein conc (y-axis).
Extrapolate the linear or quadratic (best fit) line(and equation) for Abs values of the standards.
Use the formula for std curve to calculate the protein conc of samples/unknowns.
You do this by substituting Avg Absorbance (of sample) as x into the linear/quadratic formula; y =protein conc, x = avg absorbance.
I do this easily on excel, some spectrophotometers have built-in programs.