“Quanticon” Human TNF-α (Tumor Necrosis Factor Alpha) ELISA Kit

Principle

This kit uses the Sandwich-ELISA principle. The microtiter plate strips has been pre-coated with an affinity purified antibody to Human TNF-α. Standards or samples containing Human TNF-α are added to the plate and reacted with capture antibody. A second anti-Human TNF-α antibody labelled biotin is then added and binds to Human TNF-α captured on the plate. After that, Streptavidin- Horseradish Peroxidase(SA-HRP) is added to form a sandwich complex of solid phase antibody-Human TNF-α-biotin labeled antibody-SA-HRP. And then, TMB substrate solution is added to all wells and incubated. An enzyme-catalyzed reaction generates a blue color in the solution, thereafter, stop solution is added to stop the substrate reaction and the color turns yellow. The yellow solution is read at a wavelength of 450nm. The concentration of Human TNF-α in the samples is then calculated from the OD value by establishing a standard curve.

Specification

Precision

Mean coefficient of variation for Intra-Assay and Inter-Assay: 3 samples with low, middle and high level concentration were tested for repeat multiple times, respectively.

Rate of recovery

The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.