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Human sE-Selectin-1 ELISA
Description:
Human sE-Selectin-1 ELISA
Trade Name:
Chemicon (Millipore)
Qty/Pk:
1 kit
Product Overview:
E-selectin (Endothelial Leukocyte Adhesion Molecule-1, ELAM-1) belongs to the selectin family of adhesion molecules [Hogg, 1992; Lasky, 1991]. Together with LECAM-1 (L-selectin) and GMP-140 (P-selectin), E-selectin mediates the initial interactions of leukocytes and platelets with endothelial cells [Shimizu et al., 1991; Vonandrian et al., 1992].
Molecular structure: The extracellular part of all selectins consists of an aminoterminal c-type lectin domain which specifically binds to carbohydrate ligands. This is followed by an EGF-like domain, and, in the case of E-selectin, by 6 short consensus repeats. The transmembrane portion of the molecule is followed by a short cytoplasmic tail.
Selectins provide the first, loose contacts of polymorphonuclear cells with the endothelium in areas of inflammation. The binding partner for E-selectin contains sialyl LewisX oligosaccharide or lactosaminoglycans that contain either sialic acid or fucose [Foxall, et al., 1992; Parekh, 1991]. E-selectin is expressed on cytokine-activated endothelial cells, and promotes the adhesion of leukocytes to the endothelium. This initial binding event is a prerequisite for the activation of the immune cells via inflammatory mediators [Kyan-Aung et al., 1991; Lawrence & Springer, 1991]. In contrast to GMP-140, E-selectin is maximally expressed 2-4 hours after cell activation. During the following 24-48 hours E-selectin is shed from the cytoplasmic membrane into the circulation [Pigott et al., 1992]. The circulating form of this selectin attracts neutrophils and activates the b2-integrins in preparing the cells for migration.
Soluble E-selectin levels could provide insights into several pathologies, including allergic reactions [Brasch & Sterry, 1992; Gundel et al., 1991; Mulligan et al., 1991], septic shock [Engelberts et al., 1992; Munro et al., 1991; Redl et al., 1991], vascular infection and inflammatory bowel disease [Koizumi et al., 1992].
Test Principle:
An anti-E-selectin monoclonal antibody is adsorbed onto microwells.
The pair of monoclonal antibodies used in this ELISA detect the soluble form of E-Selectin present in serum, plasma, urine, and other biological fluids.
Soluble E-selectin present in a sample or standard then binds to antibodies adsorbed to the microwells. A second, HRP-conjugated monoclonal anti-E-selectin antibody is added and binds to E-selectin captured by the first antibody.
Unbound enzyme-conjugated anti-E-selectin is removed with a wash step and HRP substrate solution is added to the wells.
An amount of colored product is formed, proportional to the amount of soluble E-selectin present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from six E-selectin standard dilutions and the E-selectin sample concentration is determined.
Application:
The E-selectin ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of soluble Endothelial Leukocyte Adhesion Molecule-1 levels in cell culture supernatants, human serum, plasma, amniotic fluid, or other body fluids. The E-selectin ELISA is for research use only. Not for use in diagnostic or therapeutic procedures.
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Applications:
The E-selectin ELISA quantitates soluble Endothelial Leukocyte Adhesion Molecule-1 levels in cell culture supernatants, human serum, plasma, amniotic fluid, or other body fluids.
Key Applications:
ELISA
Species Reactivity:
Human
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be u |