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Human VEGF ELISA Kit
Condition of sample preparation and optimal sample dilution should be determined experimentally by the investigator.
EK000013-EK0539: Human VEGF ELISA Kit
Range 31.2pg/ml-2000pg/ml (human sera, body fluids)
15.6pg/ml-1000pg/ml (cell culture supernates)
Sensitivity < 1 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of human VEGF in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF), a potent cytokine expressed by most malignant tumors, has critical roles in vasculogenesis and both physiological and pathological angiogenesis.1 VEGF produced by tumor cells potently stimulates endothelial cell proliferation and angiogenesis and plays a key role in the pathophysiology of several neoplasias.2 VEGF may also play a pivotal role in mediating the development and progression of diabetic retinopathy.3 VEGF, a major regulator of angiogenesis, binds to two receptor tyrosine kinases, KDR/Flk-1 and Flt-1.4 The VEGF gene is mapped by fluorescence in situ hybridization to chromosome 6p12 Range 31.2pg/ml-2000pg/ml (human sera, body fluids)15.6pg/ml-1000pg/ml (cell culture supernates)
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.
1. Basu, S.; Nagy, J. A.; Pal, S.; Vasile, E.; Eckelhoefer, I. A.; Bliss, V. S.; Manseau, E. J.; Dasgupta, P. S.; Dvorak, H. F.; Mukhopadhyay, D. The neurotransmitter dopamine inhibits angiogenesis induced by vascular permeability factor/vascular endothelial growth factor. Nature Med. 7: 569-574, 2001.
2. Poulaki, V.; Mitsiades, C. S.; McMullan, C.; Sykoutri, D.; Fanourakis, G.; Kotoula, V.; Tseleni-Balafouta, S.; Koutras, D. A.; Mitsiades, N. Regulation of vascular endothelial growth factor expression by insulin-like growth factor I in thyroid carcinomas. J. Clin. Endocr. Metab. 88: 5392-5398, 2003.
3. Awata, T.; Inoue, K.; Kurihara, S.; Ohkubo, T.; Watanabe, M.; Inukai, K.; Inoue, I.; Katayama, S. A common polymorphism in the 5-prime-untranslated region of the VEGF gene is associated with diabetic retinopathy in type 2 diabetes. Diabetes 51: 1635-1639, 2002.
4. Soker, S.; Takashima, S.; Miao, H. Q.; Neufeld, G.; Klagsbrun, M. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92: 735-745, 1998.
5. Wei, M.-H.; Popescu, N. C.; Lerman, M. I.; Merrill, M. J.; Zimonjic, D. B. Localization of the human vascular endothelial growth factor gene, VEGF, at chromosome 6p12. Hum. Genet. 97: 794-797, 1996.