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EK000032-EK0465: Human MMP-9 ELISA Kit
Range 156pg/ml-10,000pg/ml
Sensitivity < 5pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human MMP-9 in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
The 92-kD type IV collagenase is also known as 92-kD gelatinase, type V collagenase, gelatinase B, or matrix metalloproteinase-9 (MMP9). The 72- and 92-kDa type IV collagenases are members of a group of secreted zinc metalloproteases.1 The matrix metalloproteinases (MMPs) are able to degrade the extracellular matrix and allow angiogenesis and tumor invasion.2 Gelatinase B, a matrix metalloproteinase that has proteolytic activity against connective tissue proteins, has been suggested to be important in the connective tissue remodeling processes associated with atherogenesis and plaque rupture.3 MMP-9 is predominantly expressed in neutrophils, macrophages, and mast cells, rather than in oncogene-positive neoplastic cells.4 The polymorphism of MMP-9 acts as a genetic factor for the development of smoking-induced pulmonary emphysema.5 The standard product used in this kit is recombinant human MMP-9 with the molecular mass of 95KDa. The detected MMP-9 includes zymogen and active enzyme.
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. Collier, I. E.; Bruns, G. A. P.; Goldberg, G. I.; Gerhard, D. S. On the structure and chromosome location of the 72- and 92-kDa human type IV collagenase genes. Genomics 9: 429-434, 1991.
2. Turner, H. E.; Nagy, Z.; Esiri, M. M.; Harris, A. L.; Wass, J. A. H. Role of matrix metalloproteinase 9 in pituitary tumor behavior. J. Clin. Endocr. Metab. 85: 2931-2935, 2000.
3. Zhang, B.; Ye, S.; Herrmann, S.-M.; Eriksson, P.; de Maat, M.; Evans, A.; Arveiler, D.; Luc, G.; Cambien, F.; Hamsten, A.; Watkins, H.; Henney, A. M. Functional polymorphism in the regulatory region of gelatinase B gene in relation to severity of coronary atherosclerosis. Circulation 99: 1788-1794, 1999.
4. Coussens, L. M.; Tinkle, C. L.; Hanahan, D.; Werb, Z. MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis. Cell 103: 481-490, 2000.
5. Minematsu, N.; Nakamura, H.; Tateno, H.; Nakajima, T.; Yamaguchi, K. Genetic polymorphism in matrix metalloproteinase-9 and pulmonary emphysema. Biochem. Biophys. Res. Commun. 289: 116-119, 2001.
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