Rat A Disintegrin And Metalloproteinase With Thrombospondin 5 ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com enables the in vitro quantitative determination of rat ADAMTS5 in serum, plasma, tissue homogenates, and other biological fluids. The assay uses a pre-coated microplate, a biotinylated anti-ADAMTS5 detection antibody, and an HRP-Avidin conjugate. Following incubation with TMB substrate and an acidic stop solution, the absorbance of the resulting yellow product is read at 450 nm in a microplate reader and the ADAMTS5 level is calculated by reference to a standard curve.

Reported sensitivity is <0.064 ng/ml, the assay range is 0.156–10 ng/ml, and total assay time is 4 hours 30 minutes. In spike-and-recovery testing, average recovery was 91% in serum (range 80–102%, n=5), 91% in EDTA plasma (range 81–101%, n=5), and 84% in heparin plasma (range 80–89%, n=5). Serum is collected in serum separator tubes, allowed to clot for 30 minutes at room temperature, and centrifuged at 1000 × g for 15 minutes; plasma is collected using EDTA or heparin as anticoagulant and centrifuged at 1000 × g for 15 minutes within 30 minutes of collection; tissue is homogenized in PBS at a ratio of 1 g tissue to 9 ml PBS and centrifuged at 5000 × g for 5 minutes. Samples may be stored at +4°C for up to 5 days, at -20°C for up to 1 month, or at -80°C for up to 6 months, avoiding repeated freeze/thaw cycles.

Kit components are stored at +4°C or -20°C as instructed and include a pre-coated 96-well microplate (12 × 8 well strips), lyophilized standard, Detection Solution A, Detection Solution B, 30X Wash Buffer, Sample Dilution Buffer, TMB Substrate, Stop Solution, and plate sealers. The product is intended for research use only and is not for use in diagnostic or therapeutic procedures.