Mouse Thyrotropin Releasing Hormone ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of mouse Thyrotropin Releasing Hormone (TRH) in serum, plasma or other biological fluids. The assay uses a microtiter plate pre-coated with an antibody specific for TRH. Standards and test samples are added to the wells, where TRH present in each sample binds to the immobilized antibody. After incubation and washing, a Biotinylated Anti-TRH Antibody is added and binds the captured TRH. HRP-Avidin conjugate is then added and the plate is incubated. Following further washing, TMB substrate solution is used to visualize the HRP enzymatic reaction, producing a blue colour that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of TRH captured in each well.

Concentration of TRH is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve. Detection is colourimetric. The assay has a reported sensitivity of less than 2.77 pg/ml and a standard range of 7.81-500 pg/ml. 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.

Spike-and-recovery performance has been evaluated across sample types. In serum (n=5), average recovery was 91% with individual dilution recoveries (1:2, 1:4, 1:8, 1:16) of 87-91%, 87-107%, 74-101% and 92-97% respectively. EDTA plasma (n=5) showed an average recovery of 90% with dilution recoveries of 90-105%, 84-101%, 90-101% and 79-108%. Heparin plasma (n=5) gave an average recovery of 84% with dilution recoveries of 84-95%, 92-105%, 82-105% and 89-91%.