Human Visceral Adipose Tissue Derived Serine Protease Inhibitor ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (ELISA) from Antibodies.com is designed for the in vitro quantitative determination of human Vaspin (Visceral Adipose Tissue Derived Serine Protease Inhibitor) in serum, plasma, tissue homogenates or other biological fluids. The assay uses a 96-well microtiter plate pre-coated with an antibody specific for Vaspin. Standards and test samples are added to the wells, where Vaspin is captured by the immobilized antibody. After washing, a Biotinylated Anti-Vaspin Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate solution is then used to visualise the enzymatic reaction, producing a blue colour that changes to yellow upon addition of acidic stop solution. Absorbance is read at 450 nm in a colorimetric microplate reader, and Vaspin concentration is calculated by reference to the standard curve.

Assay performance characteristics include a sensitivity of 0.054 ng/ml and an assay range of 0.156–10 ng/ml. Total assay time is 4h 30m. Reported spike-and-recovery averages were 93% for serum (n=5, range 86–100%), 94% for EDTA plasma (n=5, range 87–102%) and 94% for heparin plasma (n=5, range 88–100%). Dilution linearity at 1:2, 1:4, 1:8 and 1:16 gave ranges of 90–99%, 82–108%, 82–96% and 92–103% for serum; 84–105%, 88–93%, 89–107% and 72–106% for EDTA plasma; and 89–92%, 88–108%, 84–102% and 90–99% for heparin plasma (n=5 each). The kit is supplied with 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 5 adhesive plate sealers, with components stored at +4°C or −20°C as indicated.