Apelin 13/Apelin (AP13/Apelin) BioAssay(TM) ELISA Kit (Human)

Catalog Number: USB-353984
Article Name: Apelin 13/Apelin (AP13/Apelin) BioAssay(TM) ELISA Kit (Human)
Biozol Catalog Number: USB-353984
Supplier Catalog Number: 353984
Alternative Catalog Number: USB-353984-48, USB-353984-96
Manufacturer: US Biological
Category: Kits/Assays
The Apelin 13/Apelin (AP13/Apelin) BioAssay(TM) ELISA Kit is a quantitative competitive assay for the detection of Apelin 13 and Apelin in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples. Detection Range: 125-8000pg/ml Sensitivity: <75pg/ml Precision: Intra-Assay CV: <8% Inter-Assay CV: <10% Assay Principle: The microtiter plate provided in this kit has been pre-coated with AP13. AP13 in the standard or sample competes with a fixed amount of plate-coated AP13 for antibody binding sites on the Antibody-Biotin reagent. Excess conjugate and unbound sample or standard are washed from the plate, after which Streptavidin-HRP (SABC) is added to each well and incubated. TMB substrate is then added to each well. The enzyme-substrate reaction is terminated by the addition of a sulfuric acid solution and the resulting color is measured spectrophotometrically at 450nm. The concentration of AP13/Apelin in the samples is determined by comparing the absorbance of the samples to the standard curve. Important Note: This kit detects both Apelin 13 and full-length Apelin. The immunogen used to generate the antibody is Apelin 13 but this antibody is 100% cross-reactive with full-length Apelin. Kit Components: *353984A: Microtiter Strips, 8x12 wells. *353984B: Standard, 2 vials 353984C: Sample/Standard Dilution Buffer, 1x20ml 353984D: Antibody-Biotin Concentrate, 1x160ul 353984E: Antibody Dilution Buffer, 1x10ml 353984F: Streptavidin-HRP Conjugate (SABC), 1x120ul 353984G: SABC Dilution Buffer, 1x10ml 353984H: TMB Substrate, 1x10ml 353984J: Stop Solution, 1x10ml 353984K: Wash Buffer, 25X, 1x30ml Storage and Stability: Store unopened *353984A at 4C, store at -20C once opened. Store unopened *353984B at 4C, once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. 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 50ul standard or sample to each well. 3. Immediately add 50ul Antibody-Biotin working solution to each well and incubate for 45 minutes at 37C 4. Aspirate and wash 3 times. 5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37C 6. Aspirate and wash 5 times. 7. Add 90ul TMB substrate. Incubate 15-20 minutes at 37C 8. Add 50ul Stop Solution. Read at 450nm immediately. 9. Calculate results.