This assay applies the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific to IL-13 has been pre-coated onto a microplate. Standards and samples are then added to the appropriate plate wells with a biotin-conjugated antibody preparation specific for IL-13 and incubated. IL-13 if present, will bind and become immobilized by the antibody pre-coated on the wells and then be "sandwiched" by biotin conjugate. Avidin is a tetramer containing four identical subunits that each has a high affinity-binding site for biotin. After washing away any unbound substances, avidin-Horseradish Peroxidase (HRP) will be added to each well and incubated. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and only those wells that contain IL-13, biotin conjugated antibody and enzyme-conjugated Avidin will exhibit a change in colour. The colour development is stopped and the intensity of the colour is measured. In order to measure the concentration of IL-13 in the samples, this package of reagent includes two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing.) According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D) versus IL-13 concentration. The concentration of IL-13 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Piattaforma
Microplate
Tipo di rilevamento
Colorimetric
Tipo di campione
Serum, plasma and cell culture supernatant.
Sensibilità
1 pg/ml
Campo di rilevamento
15.6-1000 pg/ml
Specificità
This sandwich ELISA recognises both natural and recombinant human IL-13 Ra2.
Reattività
Human
Reattività incrociata
This assay has shown no cross-reactivity with other cytokines and growth factors tested.
