Porcine Hepcidin ELISA Kit

This sandwich ELISA kit from Antibodies.com is designed for the in vitro quantitative determination of porcine Hepcidin in serum, plasma, tissue homogenates, cell lysates, cell culture supernates, or other biological fluids. The assay uses the sandwich enzyme immunoassay technique: an antibody specific for Hepcidin is pre-coated onto a 96-well microplate, standards and samples are added, and Hepcidin is captured by the immobilized antibody. After washing, wells are incubated with Biotinylated Anti-Hepcidin Antibody followed by an HRP-Avidin conjugate. TMB substrate produces a colour change proportional to Hepcidin concentration, read at 450 nm against a standard curve.

The quantitative, colourmetric assay is run on a pre-coated microplate (12 x 8 well strips) over a total assay time of 4h 30m, with a standard product range of 0.781–50 ng/ml and a sensitivity of 0.31 ng/ml. Reported average spike-and-recovery values are 92% for serum (range 84%–100%, n=5), 91% for EDTA plasma (range 83%–99%, n=5), and 93% for heparin plasma (range 85%–101%, n=5). Dilution linearity across 1:2 to 1:16 yields recoveries of 85–110% / 77–102% / 90–96% / 84–108% for serum (n=5), 88–100% / 90–104% / 76–110% / 86–93% for EDTA plasma (n=5), and 89–102% / 75–106% / 94–97% / 88–110% for heparin plasma (n=5).

Components include a pre-coated 96-well microplate (12 x 8 well strips), 2 vials of lyophilized standard, 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. Components are stored at +4°C or –20°C as instructed, and the kit should not be used past its expiration date.