Rat Hypoxia Inducible Factor 2 Alpha ELISA Kit

A sandwich enzyme-linked immunosorbent assay (ELISA) from Antibodies.com for the in vitro quantitative determination of rat HIF-2 alpha in tissue homogenates, cell lysates and other biological fluids. The assay employs a 96-well microplate pre-coated with an antibody specific for HIF-2 alpha. Standards and test samples are added to the wells, where HIF-2 alpha is captured by the immobilised antibody. After washing, a Biotinylated Anti-HIF-2 alpha Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate produces a blue product that turns yellow upon addition of acidic stop solution, and absorbance is read at 450 nm on a colorimetric microplate reader. The stated assay range is 78.1–5000 pg/ml with a sensitivity of <25 pg/ml, and the total assay time is 4 hours 30 minutes.

Validated sample matrices include serum, EDTA plasma and heparin plasma. Spike-and-recovery for serum (n=5) averaged 91% across a range of 83%–100%; EDTA plasma (n=5) averaged 96% (90%–103%); heparin plasma (n=5) averaged 94% (90%–99%). Dilution linearity was assessed at 1:2, 1:4, 1:8 and 1:16. The kit is supplied with a pre-coated 96-well microplate, lyophilised standard, Detection Solution A and B, Wash Buffer (30X), Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers. Component storage conditions vary between +4°C and -20°C, and the kit should not be used past its expiration date.