Human Sequestosome 1 ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of human SQSTM1/p62 in tissue homogenates, cell lysates or other biological fluids, and is reactive with human samples. The assay employs an antibody specific for SQSTM1/p62 pre-coated onto a 96-well microplate. Standards and test samples are added and any SQSTM1/p62 present is bound to the wells by the immobilized antibody. After washing, wells are incubated with a Biotinylated Anti-SQSTM1/p62 Antibody that binds the captured SQSTM1/p62. An HRP-Avidin conjugate is then added, followed by TMB substrate, which is catalysed by HRP to produce a blue colour that turns yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of SQSTM1/p62 captured, and concentration is calculated by reading O.D. absorbance at 450 nm against a standard curve.

The kit has a sensitivity of <0.125 ng/ml and an assay range of 0.312-20 ng/ml, with a total assay time of 4h 30m. It is read on a colorimetric microplate reader, and components are stored at +4°C or -20°C as instructed. The kit should not be used past its expiration date.

In spike-and-recovery testing (n=5), serum samples returned ranges of 80-102% (average 91%), EDTA plasma 81-101% (average 91%), and heparin plasma 80-89% (average 84%). Dilution linearity at 1:2, 1:4, 1:8 and 1:16 dilutions produced: serum 87-91%, 87-107%, 74-101%, 92-97%; EDTA plasma 90-105%, 84-101%, 90-101%, 79-108%; and heparin plasma 84-95%, 92-105%, 82-105%, 89-91%. The kit is suitable for cell lysates and tissue homogenates and includes a pre-coated microplate, lyophilized standard, detection solutions A and B, wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers.