This ELISA kit applies to the in vitro quantitative determination of Human CRH concentrations in serum, plasma and other biological fluids.

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Human CRH. During the reaction, Human CRH in the sample or standard competes with a fixed amount of Human CRH on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Human CRH. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Human CRH in the samples is then determined by comparing the OD of the samples to the standard curve.

This kit recognizes Human CRH in samples. No significant cross-reactivity or interference between Human CRH and analogues was observed.

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CRH were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CRH were tested on 3 different plates, 20 replicates in each plate.

The recovery of Human CRH spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Samples were spiked with high concentrations of Human CRH and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

An unopened kit can be stored at 2-8°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).