Rat Midkine ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of rat Midkine in serum, plasma or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, using an antibody specific for Midkine that is pre-coated onto a 96-well microtiter plate. Standards and test samples are added to the wells, where Midkine is bound by the immobilized antibody. After incubation and washing, a Biotinylated Anti-Midkine Antibody binds the captured Midkine in each well. HRP-Avidin conjugate is then added, and following further incubation and washing, a TMB substrate solution is used to visualize the enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of Midkine captured, and the concentration is calculated by reading the optical density at 450 nm on a colorimetric microplate reader against the standard curve.

The assay reports a sensitivity of 7.2 pg/ml and a range of 15.6-1000 pg/ml, with a total assay time of approximately 4 hours 30 minutes. Validation data report recovery in serum (n=5) ranging from 80% to 102% with an average of 91%, in EDTA plasma (n=5) ranging from 81% to 99% averaging 90%, and in heparin plasma (n=5) ranging from 80% to 89% averaging 84%. Dilution linearity is reported across serum, EDTA plasma, and heparin plasma at 1:2, 1:4, 1:8, and 1:16 dilutions.

The kit is supplied with a pre-coated 96-well microplate arranged as 12 x 8 well strips, lyophilized standard, detection solution A, detection solution B, wash buffer (30X), sample dilution buffer, TMB substrate, stop solution, and plate sealers. Kit components are stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.