Human Gonadotropin Releasing Hormone ELISA Kit

From Antibodies.com, this ELISA kit enables the in vitro quantitative determination of human Gonadotropin Releasing Hormone (GnRH). It employs the sandwich enzyme immunoassay technique, using a GnRH-specific antibody pre-coated onto a 96-well microplate. Standards and test samples are added to the wells, and GnRH present in each sample is bound by the immobilised antibody. After washing, a Biotinylated Anti-GnRH Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of the acidic stop solution. Absorbance is read at 450 nm in a microplate reader and GnRH concentration is calculated by reference to the standard curve.

The kit is suitable for serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Reported sensitivity is <0.064 ng/ml, with a measurable range of 0.156–10 ng/ml. Detection is colourimetric on a standard microplate reader platform, and the total assay time is 4 hours 30 minutes. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.

Recovery has been characterised across matrices. In serum (n=5), recovery ranged from 80% to 102% with an average of 91%. EDTA plasma (n=5) showed 81%–100% recovery (average 90%), while heparin plasma (n=5) gave 80%–89% (average 84%). Dilution linearity was also assessed: serum (n=5) at 1:2, 1:4, 1:8 and 1:16 dilutions yielded 87–91%, 87–107%, 74–101% and 92–97% respectively; EDTA plasma (n=5) gave 90–105%, 84–101%, 90–101% and 79–108%; and heparin plasma (n=5) gave 84–95%, 92–105%, 82–105% and 89–91%.