The Phospho Tau (P181) BioAssay(TM) ELISA Kit is a quantitative sandwich assay for the detection of Phospho Tau (P181) in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples. Detection Range: 7.813-500pg/ml Sensitivity: 4.688pg/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 Phospho Tau (P181). Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to Phospho Tau (P181). 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 Phospho Tau (P181), 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 Phospho Tau (P181) in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *519426A: Microtiter Strips, 1x96 wells *519426B: Standard, 2x1 vial 519426C: Sample/Standard Dilution Buffer, 1x20ml 519426D: Antibody (Biotin) (Concentrated), 1x120ul 519426E: Antibody Dilution Buffer, 1x10ml 519426F: Streptavidin (HRP) (SABC), 1x120ul 519426G: SABC Dilution Buffer, 1x10ml 519426H: TMB Substrate, 1x10ml 519426J: Stop Solution, 1x10ml 519426K: Wash Buffer, 25X, 1x30ml Storage and Stability: Store unopened *519426B at 4C, store at -20C once opened. Store unopened *519426B 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 and samples to wells! 2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37C. Aspirate and wash 2 times. 3. Add 100ul Antibody (Biotin) working solution to each well and incubate for 60 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 10-20 minutes at 37C 8. Add 50ul Stop Solution. Read at 450nm immediately. 9. Calculate results
