Rat Hypoxia Inducible Factor 1 Alpha ELISA Kit

A sandwich ELISA kit from Antibodies.com for the in vitro quantitative determination of rat HIF-1 alpha in tissue homogenates, cell lysates, cell culture supernates and other biological fluids. The assay uses the sandwich enzyme immunoassay technique on a 96-well microplate pre-coated with antibody specific for HIF-1 alpha. Standards and samples are added to the wells, followed by a biotinylated anti-HIF-1 alpha antibody and an HRP-avidin conjugate. After incubation and washing, TMB substrate is added; the resulting blue product turns yellow upon addition of acidic stop solution. The intensity of the yellow colour, read at 450 nm on a colorimetric microplate reader, is proportional to HIF-1 alpha concentration and is calculated against a standard curve.

Quantitative performance includes an assay range of 0.156–10 ng/ml and a sensitivity of <0.057 ng/ml, with a total assay time of 4 h 30 min. The kit is supplied with a pre-coated 96-well microplate (12 × 8-well strips), lyophilised standard, detection solutions A and B, 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 not used past the kit's expiration date.

Spike-and-recovery testing in serum, EDTA plasma and heparin plasma produced average recoveries of 91 %, 96 % and 95 % respectively, across dilution factors from 1:2 to 1:16, demonstrating reliable quantification across common biological fluid types. The HIF-1 alpha target is also known by synonyms including ARNT-interacting protein, HIF1A, Hypoxia-inducible factor 1-alpha, MOP1 and PASD8.