Human Endothelin-1
The Endothelin-1 (ET-1) Enzyme Immunometric Assay (EIA) kit is a complete kit for the quantitative determination of ET-1 in plasma, serum, and culture fluids.
The Endothelin-1 (ET-1) Enzyme Immunometric Assay (EIA) kit is a complete kit for the quantitative determination of ET-1 in plasma, serum, and culture fluids. ET-1 has identical amino acid sequence for human, mouse, rat, cow, dog, pig, and rabbit. Please read the complete kit insert before performing this assay.
Endothelins (ET) were first identified as endothelin-derived relaxation factors then again as an endothelin-derived contracting factor. In 1988, the isolation and sequence of a 21 amino acid peptide was reported and identified by the more common name endothelin 1. Three distinct endothelin genes encode unique but highly related peptides, ET-1, ET-2, and ET-31. The predominant physical feature of ET’s is the central helical core that is stabilized and includes two intra-chain disulfide bonds. This conformation is necessary for high affinity binding by the ETA receptor, but is not for binding by ETB2. Translated as a pre-pro peptide and translocated into circulation as the pro form Big ET-1, bioactive ET-1 is released when Big ET-1 is cleaved by endothelin-converting enzymes at the site of action. Once released, ET-1 is able to elicit different vaso-actions depending on which receptor is bound3. The basal circulating level of ET-1 is reported to be < 1 to 3 pg/mL but is known to be elevated in atrial and pulmonary hypertensions, atherosclerosis, congestive heart failure, cancer4 and variably related to lung diseases such as COPD and asthma5.
Samples and standards are added to wells coated with a monoclonal antibody specific for ET-1. The plate is then incubated.
The plate is washed, leaving only bound ET-1 on the plate. A solution of HRP labeled monoclonal antibody to ET-1 is then added. This binds the ET-1 captured on the plate. The plate is then incubated.
The plate is washed to remove excess HRP labeled antibody. TMB Substrate solution is added. The substrate generates a blue color when catalyzed by the HRP.
Stop solution is added to stop the substrate reaction. The resulting yellow color is read at 450 nm. The amount of signal is directly proportional to the level of ET-1 in the sample.
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