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EK000043-EK0519: Human TIE2 ELISA Kit
Range 156pg/ml-10, 000pg/ml
Sensitivity < 5 pg/ml
Specificity: cross-reactivates with human TIE1< 0.1%.
Application: for quantitative detection of human TIE2 in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Tyrosine kinase with Ig and EGF homology domain 2 (Tie-2), also called TEK tyrosine kinase, endothelial (TEK). Tie-2 and tie-1 are expressed in early embryonic vascular system and in maternal decidual vascular endothelial cells, where the vasculature undergoes an active angiogenesis. Tie-2, but not tie-1, expression was also detected in extraembryonic mesoderm of the amnion.1 Angiogenesis is coordinated with follicular cell growth in goitrogenesis. The angiopoietins, Ang-1 and Ang-2, are angiogenic growth factors acting through Tie-2. Tie-2 and Ang-1 are expressed in thyroid epithelial and endothelial cells, and Tie-2 is regulated by TSH and cAMP in follicular cells. And Tie-2 expression is increased in goiter in both humans and rats, consistent with a role in goitrogenesis.2 Tie2/Ang-1 signaling pathway plays a critical role in the maintenance of HSCs in a quiescent state in the BM niche.3 And the Tie-2 signaling pathway is also critical for endothelial cell-smooth muscle cell communication in venous morphogenesis.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. Sato, T. N.; Qin, Y.; Kozak, C. A.; Audus, K. L. : tie-1 and tie-2 define another class of putative receptor tyrosine kinase genes expressed in early embryonic vascular system. Proc. Nat. Acad. Sci. 90: 9355-9358, 1993.
2. Ramsden, J. D.; Cocks, H. C.; Shams, M.; Nijjar, S.; Watkinson, J. C.; Sheppard, M. C.; Ahmed, A.; Eggo, M. C. : Tie-2 is expressed on thyroid follicular cells, is increased in goiter, and is regulated by thyrotropin through cyclic adenosine 3-prime,5-prime-monophosphate. J. Clin. Endocr. Metab. 86: 2709-2716, 2001.
3. Arai, F.; Hirao, A.; Ohmura, M.; Sato, H.; Matsuoka, S.; Takubo, K.; Ito, K.; Koh, G. Y.; Suda, T. : Tie2/Angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell 118: 149-161, 2004.
4. Vikkula, M.; Boon, L. M.; Carraway, K. L., III; Calvert, J. T.; Diamonti, A. J.; Goumnerov, B.; Pasyk, K. A.; Marchuk, D. A.; Warman, M. L.; Cantley, L. C.; Mulliken, J. B.; Olsen, B. R. : Vascular dysmorphogenesis caused by an activating mutation in the receptor tyrosine kinase TIE2. Cell 87: 1181-1190, 1996.
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