Assay Principle

This kit employs double antibody sandwich ELISA technology: Capture Antibody is coated on the microplate to capture IFN-γ from samples and standards. After washing, biotin-labeled Detection Antibody is added and incubated, followed by washing to form a "Capture Antibody-Antigen-Detection Antibody" immune-complex. Subsequently, Streptavidin-Horseradish Peroxidase (SA-HRP) is added and incubated. After incubation and washing, TMB Substrate is added for color development. If the target analyte is present in the sample, a blue color develops. Stop Solution is then added to terminate the reaction. During the assay, unbound components are washed away. The Optical Density (OD) is measured at 450 nm using a microplate reader. The intensity of the color is proportional to the IFN-γ concentration in the sample, and the concentration is calculated by plotting a standard curve.

Recovery

Recovery was tested by spiking known concentrations of Human IFN-γ into different sample matrices. The recovery range and average recovery are shown below.

Sensitivity

The minimum detectable concentration (sensitivity) of Human IFN-γ, determined by testing samples, is 0.78pg/mL.