Assay Principle: 

The DetectX® Endothelin-1 (ET-1) ELISA Kit quantitatively measures ET-1 levels in serum, plasma, and tissue culture media. This sandwich ELISA has a total run time of 2.5 hours, facilitating efficient and accurate analysis. Please read the complete kit insert for complete instructions before beginning the assay. The kit includes an Endothelin-1 standard to create an accurate standard curve.

Protocol Summary:

• Introduce standards or diluted samples into the provided transparent microtiter plate coated with human Endothelin-1 antibody.
• Incubate the plate covered at room temperature for 1 hour.
• Add peroxidase-conjugated human Endothelin-1 antibody and incubate for another hour.
• After incubation, wash away the excess conjugated antibody and add the TMB substrate. The TMB substrate reacts with the bound antibody to generate a detectable signal at 450nm.
• Calculate the concentration of Endothelin-1 using the signal intensity and standard curve.

Background:

Endothelin-1 (ET-1), a peptide of 21 amino acid residues, is a pleiotropic molecule known for its action as a potent vasoconstrictor. It is part of the Endothelin family, distinguished by two essential disulfide bridges and six conserved amino acids at the C-terminus. The initial synthesis of Human ET-1 is a pre-pro-polypeptide of 212 amino acids. A signal peptidase proteolytically cleaves the pre-pro-polypeptide producing pro-ET-1. A Furin-like protease further processes pro-ET-1 to yield Big ET-1. The vascular endothelium is a primary source of ET-1, but it is also expressed in various other cell types, including leukocytes, smooth muscle cells, and astrocytes.

ET-1 production can be stimulated by mechanical forces, hormones, and pro-inflammatory cytokines, while its production is inhibited by factors like nitric oxide (NO) and atrial natriuretic peptide (ANP). Due to its significant role in vascular tone and blood pressure regulation, ET-1 is a major focus cardiovascular research. It has implications in conditions such as hypertension, heart failure, and other cardiovascular diseases. The DetectX® Endothelin-1 ELISA Kit enables researchers and clinicians to accurately measure ET-1 levels, providing valuable insights into cardiovascular health and disease mechanisms.