Rat 3-Beta-Hydroxysteroid Dehydrogenase Type 1 ELISA Kit

From Antibodies.com, this sandwich ELISA enables the in vitro quantitative determination of rat 3-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD3B1) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.

An antibody specific for HSD3B1 is pre-coated onto a 96-well microplate (12 x 8 well strips). Standards and test samples are added to the wells and HSD3B1 present in each sample is bound by the immobilised antibody. After washing, a biotinylated anti-HSD3B1 antibody is added, followed by an HRP-avidin conjugate. TMB substrate visualises the enzymatic reaction, producing a blue colour that changes to yellow on addition of stop solution. Absorbance is read at 450 nm in a microplate reader and HSD3B1 concentration is calculated by reference to the standard curve.

The working range is 0.156–10 ng/ml with a sensitivity of <0.063 ng/ml, and the total assay time is 4 h 30 min. Reported recovery is 83–99% in serum (average 91%, n = 5), 80–100% in EDTA plasma (average 90%, n = 5) and 82–99% in heparin plasma (average 90%, n = 5). Dilution linearity was assessed at 1:2, 1:4, 1:8 and 1:16 dilutions. Across these, serum recovery (n = 5) ranged 74–109%, EDTA plasma 78–110% and heparin plasma 77–109%. Kit components are stored at +4 °C or -20 °C as instructed and the kit should not be used past its stated expiry date.