Interleukin 35 (IL-35) BioAssay(TM) ELISA Kit (Human)

The IL-35 (Interleukin 35) BioAssay(TM) ELISA Kit is a quantitative sandwich assay for the detection of IL-35 in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples. Range: 15.625-1000pg/ml, Sensitivity: <9.375pg/ml, Precision: Intra-assay CV: <8%, Inter-assay CV: <10%, Assay Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to IL-35. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to IL-35. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain IL-35, biotin-conjugated antibody and enzyme-conjugated streptavidin complex will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of IL-35 in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *356731A: Microtiter Strips, 1x96 wells (8x12 wells)., *356731B: Standard, 2x1 vial, 356731C: Sample/Standard Dilution Buffer, 1x20ml, 356731D: Antibody (Biotin) (Concentrated), 1x120ul, 356731E: Antibody Dilution Buffer, 1x10ml, 356731F: Streptavidin (HRP) (SABC), 1x120ul, 356731G: SABC Dilution Buffer, 1x10ml, 356731H: TMB Substrate, 1x10ml, 356731J: Stop Solution, 1x10ml, 356731K: Wash Buffer, 25X, 1x30ml, Storage and Stability: Store unopened *356731A at 4ºC, store at -20ºC once opened. Store unopened *356731B at 4°C, once reconstituted, store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap. Assay Summary: 1. Wash plate 2 times before adding standards, samples and control (zero) to wells!, 2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C. Aspirate and wash 2 times., 3. Add 100ul Antibody (Biotin) working solution to each well and incubate for 60 minutes at 37°C, 4. Aspirate and wash 3 times., 5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C, 6. Aspirate and wash 5 times., 7. Add 90ul TMB substrate. Incubate 15-30 minutes at 37°C, 8. Add 50ul Stop Solution. Read at 450nm immediately., 9. Calculate results