Rat Histamine Receptor H4 ELISA Kit

Antibodies.com provides a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of rat Histamine Receptor H4 (H4R) in tissue homogenates, cell lysates or other biological fluids. The kit employs the sandwich enzyme immunoassay technique in which an antibody specific for H4R is pre-coated onto a microplate. Standards and samples are added to the wells, where H4R present in each sample is captured by the immobilized antibody. After incubation and washing, a Biotinylated Anti-H4R Antibody is added, followed by an HRP-Avidin conjugate and further incubation. TMB substrate solution is then used to visualise the HRP enzymatic reaction, producing a blue product that changes to yellow upon addition of acidic stop solution. The absorbance is measured at 450 nm in a colorimetric microplate reader and the H4R concentration is calculated by reference to the standard curve.

The kit offers a sensitivity of <0.055 ng/ml and an assay range of 0.156–10 ng/ml, with a total assay time of 4h 30m. The platform is a pre-coated 12 x 8 well strip microplate. Spike-and-recovery performance is reported as: Serum (n=5) 80%–102% (average 91%), EDTA Plasma (n=5) 81%–100% (average 90%), Heparin Plasma (n=5) 80%–89% (average 84%). Dilution linearity across 1:2, 1:4, 1:8, and 1:16 dilutions is provided, for example Serum (n=5) 87–91%, 87–107%, 74–101%, 92–97%; EDTA Plasma (n=5) 90–105%, 84–101%, 90–101%, 79–108%; Heparin Plasma (n=5) 84–95%, 92–105%, 82–105%, 89–91%.

Kit components include a pre-coated 96-well microplate (12 x 8 well strips), lyophilized standard (2 vials), Detection Solution A (120 μl), Detection Solution B (120 µl), Wash Buffer 30X (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml), and plate sealers (5 adhesive strips). Components should be stored at +4°C or -20°C as instructed and the kit must not be used past its expiration date.