Assay Principle

This kit employs double antibody sandwich ELISA technology: Capture Antibody is coated on the microplate to capture TNF-α 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 TNF-α concentration in the sample, and the concentration is calculated by plotting a standard curve.

Typical Data

The following data and curve are for reference only. The experimenter must establish a standard curve based on their own experimental data.

Sensitivity

The minimum detectable concentration (sensitivity) of Human TNF-α, determined by testing samples, is 7.81pg/mL.