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EK000072-EK0501: Human E-Selectin ELISA Kit
Range 62.5pg/ml-4000pg/ml
Sensitivity < 4 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of human E-Selectin in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
E-selectin, also called endothelial-leukocyte adhesion molecule-1 (ELAM-1), is a cell surface glycoprotein expressed by cytokine-activated endothelium that mediates the adhesion of blood neutrophils.1 An increased expression of E-selectin has been observed in the arterial endothelium interacting with lymphocytes and macrophages in human atherosclerotic lesions.2 E-selectin plays a critical role in mediating tissue-specific homing of T cells into skin, and of primitive hematopoietic progenitor cells (HPCs) into bone marrow (BM).3 A structurally and functionally related group of genes, lymph node homing receptor (LHR), granule membrane protein 140 (GMP-140), and endothelial leukocyte adhesion molecule 1 (ELAM-1) are shown to constitute a gene cluster on mouse and human chromosome 1.4 The standard product used in this kit is recombinant human E-Selectin, excluding intercellular E-Selectin and transmembrane domain. It consists of totally 535 amino acids with the molecular mass of 58.6KDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Collins, T.; Williams, A.; Johnston, G. I.; Kim, J.; Eddy, R.; Shows, T.; Gimbrone, M. A., Jr.; Bevilacqua, M. P. Structure and chromosomal location of the gene for endothelial-leukocyte adhesion molecule 1. J. Biol. Chem. 266: 2466-2473, 1991.
2. Zheng, F.; Chevalier, J. A.; Zhang, L. Q.; Virgil, D.; Ye, S. Q.; Kwiterovich, P. O. An HphI polymorphism in the E-selectin gene is associated with premature coronary artery disease. Clin. Genet. 59: 58-64, 2001.
3. Dimitroff, CJ.; Lee, JY.; Rafii, S.; Fuhlbrigge, RC.; Sackstein, R. CD44 is a major E-selectin ligand on human hematopoietic progenitor cells. J Cell Biol. 2001 Jun 11;153(6):1277-86.
4. Watson, M. L.; Kingsmore, S. F.; Johnston, G. I.; Siegelman, M. H.; Le Beau, M. M.; Lemons, R. S.; Bora, N. S.; Howard, T. A.; Weissman, I. L.; McEver, R. P.; Seldin, M. F. Genomic organization of the selectin family of leukocyte adhesion molecules on human and mouse chromosome 1. J. Exp. Med. 172: 263-272, 1990.
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