One antibody molecule can be labeled with multiple biotin molecules,
and one biotin molecule can bind a horseradish peroxidase labeled
avidin, thus bringing a large amount of horseradish peroxidase bound
to the antibody, which has higher sensitivity and signal amplification
effect than the traditional direct horseradish peroxidase labeled
antibody.
Intended use
This immunoassay kit allows for the in vitro quantitative determination of human Catechol O-methyltransferase ,TNF-alpha concentrations in serum, Plasma , tissue homogenates and Cell culture supernates and Other biological fluids.
Principle
The microtiter plate provided in this kit has been pre-coated with an antibody specific to TNF-alpha. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for TNF-alpha and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain TNF-alpha, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of TNF-alpha in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Assay range
15.6-1000pg/ml
Intra-Assay: CV≤ 5.2%
Inter-Assay: CV≤ 7.4%
Recovery: 70 - 110 percent
Sensitivity
