This assay applies the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific to IL-13 Ra2 has been pre-coated onto a microplate. Standards and samples are then added to the appropriate plate wells with a biotin-conjugated polyclonal antibody preparation specific for IL-13 Ra2 and incubated. IL-13 Ra2 if present, will bind and become immobilized by the antibody pre-coated on the wells and then be "sandwiched" by biotin conjugate. After washing away any unbound substances. In order to quantitate the amount of IL-13 Ra2 present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) 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 Ra2, biotin conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The color development is stopped and the intensity of the color is measured.
In order to measure the concentration of IL-13 Ra2 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 Ra2 concentration (ng/ml). The concentration of IL-13 Ra2 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Sensitivity: <10ng/ml
Assay Range: 10-500ng/ml
Kit Components:I8435-09B1. IL-13 Ra2 Microtiter Plate 1x96 wellsI8435-09B2. Biotin Conjugate, 1x6mlI8435-09B3. Avidin Conjugate, 1x15ml I8435-09B4. IL-13 Ra2 STANDARD 2x0.5ug I8435-09B5. CALIBRATOR DILUENT I, 1x25ml For serum/plasma testing
I8435-09B6. CALIBRATOR DILUENT II, 1x25ml For cell culture supernatant testing
Storage and Stability:Store components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
