Porcine CRH (Corticotropin Releasing Hormone) ELISA Kit

This ELISA kit from Assay Genie is designed for the quantitative detection of Corticotropin-Releasing Hormone (CRH) in porcine samples. The assay is reactive with porcine (swine) samples and is suitable for use with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Detection is performed using a colorimetric method employing a TMB substrate, with absorbance measured at 450 nm on a microplate reader. Reported sensitivity is 0.938 pg/mL with a measurable range of 1.563-100 pg/mL.

The assay procedure involves equilibration of the TMB substrate, preparation of standards and samples (minimum 1:2 dilution), binding with a biotin-labeled antibody, washing, color development, and a stop reaction. Reported linearity across dilutions of 1:2, 1:4, and 1:8 spans 81-105% across serum, EDTA plasma, and heparin plasma. Recovery ranges from 87-104% with averages of 96% (serum), 97% (EDTA plasma), and 98% (heparin plasma). The kit is supplied with a dismountable ELISA microplate, lyophilized standard, biotin-labeled antibody (100X), HRP-Streptavidin conjugate (SABC, 100X), TMB substrate, sample dilution buffer, antibody dilution buffer, SABC dilution buffer, stop solution, wash buffer (25X), plate sealer, and a technical manual. Storage is recommended at 2-8°C (sealed), and cryopreservation should be avoided.

Sample preparation protocols include allowing blood to clot and centrifuging serum at 1000 × g for 20 minutes, collecting plasma with EDTA anticoagulant and centrifuging at 1000 × g for 15 minutes at 2-8°C, centrifuging cell culture supernatant at 1000 × g for 20 minutes at 4°C, lysing cells and centrifuging at 10,000 rpm for 10 minutes for cell lysates, and homogenising tissue in PBS with protease inhibitors followed by centrifugation at 5000 × g for 5 minutes. CRH is described as a key neuroendocrine peptide involved in the regulation of the stress response and the hypothalamic-pituitary-adrenal axis, and accurate measurement of CRH in porcine samples is positioned as a tool for research into stress-related and hormonal disorders in pigs.