Rat Follicle Stimulating Hormone ELISA Kit

From Antibodies.com, this competitive ELISA (cELISA) kit is designed for the in vitro quantitative determination of rat FSH beta in serum, plasma or other biological fluids. The assay uses the competitive enzyme immunoassay technique on a pre-coated 96-well microplate arranged as 12 x 8 well strips, where FSH beta antigen is immobilised. During incubation, FSH beta present in samples or standards competes with the fixed antigen for binding sites on a Biotinylated Anti-FSH beta Antibody. After washing to remove unbound antibody, an HRP-Avidin conjugate is added, followed by TMB substrate solution that produces a blue colour through enzymatic catalysis; the reaction turns yellow upon addition of acidic stop solution. The intensity of yellow is inversely proportional to the FSH beta concentration, which is quantified by reading absorbance at 450 nm in a colorimetric microplate reader and referring to a standard curve.

The assay has a range of 0.468–30 mIU/ml and a sensitivity of <0.21 mIU/ml, with a total assay time of 2h 30m. Supplied components include the pre-coated 96-well microplate, 2 vials of lyophilised standard, 70 μl of Detection Solution A, 120 μl of Detection Solution B, 20 ml of 30X wash buffer, 45 ml of sample dilution buffer, 9 ml of TMB substrate, 6 ml of stop solution, 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. Reported average recovery from serum (n=5) is 91% (range 80–102%), from EDTA plasma (n=5) is 90% (range 81–100%), and from heparin plasma (n=5) is 84% (range 80–89%).