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EK000025-EK0329: Human Eotaxin ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 2 pg/ml
Specificity: cross-reactivates with MCP-2, MCP-3 <2.5%, and with MCP-1 <0.25%.
Application: for quantitive detection of human Eotaxin in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Eotaxin, also known as CCL11, is a potent inducer of eosinophil chemotaxis and is considered as a selective ligand of the CC chemokine receptor 3 (CCR3), which is expressed on eosinophils, basophils, and Th2 lymphocytes.1 Eotaxin is assumed to be involved in eosinophilic inflammatory diseases such as atopic dermatitis, allergic rhinitis, asthma and parasitic infections.2 The gene maps to chromosome 17 and is expressed constitutively at high levels in small intestine and colon, and at lower levels in various other tissues. The deduced mature protein sequence is 66% identical to human monocyte chemoattractant protein-1, and 60% identical to guinea pig eotaxin. Recombinant human eotaxin produced in insect cells induces a calcium flux response in normal human eosinophils, but not in neutrophils or monocytes.3 The human eotaxin gene is cloned and found to be 61.8 and 63.2% identical at the amino acid level to guinea pig and mouse eotaxin.4
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. Ogilvie, P.; Bardi, G.; Clark-Lewis, I.; Baggiolini, M.; Uguccioni, M. Eotaxin is a natural antagonist for CCR2 and an agonist for CCR5. Blood 97: 1920-1924, 2001.
2. Hein, H.; Schluter, C.; Kulke, R.; Christophers, E.; Schroder, J.-M.; Bartels, J. Genomic organization, sequence, and transcriptional regulation of the human eotaxin gene. Biochem. Biophys. Res. Commun. 237: 537-542, 1997.
3. Kitaura, M.; Nakajima, T.; Imai, T.; Harada, S.; Combadiere, C.; Tiffany, H. L.; Murphy, P. M.; Yoshie, O. Molecular cloning of human eotaxin, an eosinophil-selective CC chemokine, and identification of a specific eosinophil eotaxin receptor, CC chemokine receptor 3. J. Biol. Chem. 271: 7725-7730, 1996.
4. Ponath, P. D.; Qin, S.; Ringler, D. J.; Clark-Lewis, I.; Wang, J.; Kassam, N.; Smith, H.; Shi, X.; Gonzalo, J.-A.; Newman, W.; Gutierrez-Ramos, J.-C.; Mackay, C. R. Cloning of the human eosinophil chemoattractant, eotaxin: expression, receptor binding, and functional properties suggest a mechanism for the selective recruitment of eosinophils. J. Clin. Invest. 97: 604-612, 1996.
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