The P62/SQSTM1 BioAssay™ ELISA Kit is a quantitative sandwich assay for the detection of P62/SQSTM1 in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples.
Detection Range:0.156-10ng/ml
Sensitivity:0.094ng/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 P62/SQSTM1. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to P62/SQSTM1. 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 P62/SQSTM1, 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 P62/SQSTM1 in the sample is then determined by comparing the O.D. of the sample to the standard curve.
Storage and Stability:Store unopened *519410A at 4ºC; store at -20ºC once opened. Store unopened *519410B 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 and samples 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°C6. Aspirate and wash 5 times
7. Add 90ul TMB substrate. Incubate 10-20 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately
