Human Endothelin 2 ELISA Kit

This competitive Enzyme-Linked Immunosorbent Assay (cELISA) from Antibodies.com is designed for the in vitro quantitative determination of human Endothelin 2/ET-2 in serum, plasma or other biological fluids. The 96-well microplate is pre-coated with Endothelin 2/ET-2 antigen, and the assay employs a competitive immunoassay technique in which Endothelin 2/ET-2 present in samples or standards competes with the fixed amount of immobilized antigen for binding sites on a biotinylated anti-Endothelin 2/ET-2 antibody. Following incubation and washing, an HRP-Avidin conjugate is added and TMB substrate solution is used to visualise the enzymatic reaction, producing a blue colour that changes to yellow after addition of acidic stop solution. Absorbance is read at 450 nm on a colorimetric microplate reader; the density of the yellow colour is inversely proportional to the Endothelin 2/ET-2 concentration, which is then calculated against a standard curve.

The kit has a sensitivity of <11.6 pg/ml and an assay range of 31.2-2000 pg/ml, with a total assay time of 2h 30m on a pre-coated microplate (12 x 8 well strips). Recovery was evaluated in serum (range 80%-102%, average 91%, n=5), EDTA plasma (range 81%-100%, average 90%, n=5) and heparin plasma (range 80%-89%, average 84%, n=5). Dilution linearity was assessed at 1:2, 1:4, 1:8 and 1:16 dilutions across the same sample types, with serum recoveries of 87-91%, 87-107%, 74-101% and 92-97% respectively, EDTA plasma recoveries of 90-105%, 84-101%, 90-101% and 79-108% respectively, and heparin plasma recoveries of 84-95%, 92-105%, 82-105% and 89-91% respectively.

Kit components include a pre-coated 96 well microplate (12 x 8 well strips), lyophilized standard, Detection Solution A, Detection Solution B, 30X Wash Buffer, Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers. Components should be stored at +4°C or -20°C as instructed, and the kit must not be used past its expiration date.