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Specifications
- Assay Type Sandwich ELISA
- Sample Types Serum, Plasma, Tissue Culture Media
- Sensitivity 0.579 pg/mL
- Species Human, Bovine, Porcine, Dog, Rat, Mouse
- Assay Duration 2.5 Hours
- Samples/Plate 39 in Duplicate
- Readout Colorimetric, 450 nm
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Standard Curve
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Description
Assay Principle:
The Endothelin-1 (ET-1) ELISA Kit quantitatively measures Endothelin-1 (ET-1) levels in serum, plasma, and tissue culture media. The Endothelin-1 (ET-1) ELISA Kit is a sandwich ELISA with a run time of 2.5 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided Endothelin-1 standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our human Endothelin-1 antibody and incubate covered at room temperature for 1 hour. Add the peroxidase-conjugated human Endothelin-1 antibody. Then incubate the mixture covered at room temperature for 1 hour. The immunological reaction occurs between the peroxidase-conjugated Endothelin-1 antibody and the Endothelin-1 antigen in the sample or standard.
After the 1-hour incubation, wash away the excess peroxidase-conjugated Endothelin-1 antibody and add the TMB substrate. The TMB substrate reacts with the bound peroxidase-conjugated Endothelin-1 antibody generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the Endothelin-1 concentration in the samples.
Background:
Endothelin-1 (ET-1), a peptide of 21 amino acid residues, is a pleiotropic molecule known for its action as a potent vasoconstrictor. ET-1 is one of a family of three proteins encoded by distinct genes. All members of the Endothelin family contain two essential disulfide bridges and six conserved amino acid residues 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 an abundant source of ET-1. Leukocytes, smooth muscle cells, mesangial cells, cardiac myocytes, and astrocytes can also express ET-1. Endothelial cells can induce ET-1 by mechanical stimulation, hormones, and pro-inflammatory cytokines. Nitric oxide (NO), cyclic nucleotides, prostacyclin, and atrial natriuretic peptide (ANP) inhibit the production of ET-1.