Human A Disintegrin And Metalloproteinase With Thrombospondin 7 ELISA Kit

This Antibodies.com assay kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) intended for the in vitro quantitative determination of human ADAMTS7 in tissue homogenates, cell lysates or other biological fluids, with reactivity confirmed for human samples.

The kit employs the sandwich enzyme immunoassay technique. An antibody specific for ADAMTS7 is pre-coated onto a 96-well microplate. Standards and test samples are added to the wells, where ADAMTS7 present in each sample is bound by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-ADAMTS7 Antibody is added, which binds the captured ADAMTS7 in each well. HRP-Avidin conjugate is then added and the plate is incubated. TMB substrate solution is used to visualise the HRP enzymatic reaction, producing a blue colour that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of ADAMTS7 captured, and concentration is calculated by reading O.D. absorbance at 450 nm against a standard curve.

Reported specifications include a product range of 0.156–10 ng/ml and a sensitivity of 0.059 ng/ml. The platform is a pre-coated microplate arranged as 12 x 8 well strips, with an assay time of 4h 30m, read on a colorimetric microplate reader. Recovery (n=5) was 80%–102% (average 91%) for serum, 81%–101% (average 91%) for EDTA plasma, and 80%–89% (average 84%) for heparin plasma. Dilution linearity across 1:2, 1:4, 1:8 and 1:16 dilutions spanned 87%–91%, 87%–107%, 74%–101% and 92%–97% for serum; 90%–105%, 84%–101%, 90%–101% and 79%–108% for EDTA plasma; and 84%–95%, 92%–105%, 82%–105% and 89%–91% for heparin plasma. Kit components are a pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard (2 vials), Detection Solution A (120 μl), Detection Solution B (120 µl), 30X Wash Buffer (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml) and plate sealers (5 adhesive strips), with storage temperatures of -20°C for most components and +4°C for the wash buffer, TMB substrate and stop solution. The kit is cited in 1 publication.