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EK000054-EK0462: Mouse MMP-3 ELISA Kit
Range 156pg/ml-10,000pg/ml
Sensitivity < 10pg/ml
Specificity: cross-reactivates with MMP-10 approximately 2%, and no detectable cross-reactivity with other MMPs.
Application: for quantitative detection of mouse MMP-3 in sera, plasma, body fluids, tissue lysates or cell culture supernatant.
Expiration: four months at 4°C and eight months at -20°C.
Background
Matrix metalloproteinase-3 (MMP-3) also called stromelysin or transin, is a proteoglycanase closely related to collagenase (MMP1) with a wide range of substrate specificities. The complete primary structure for human MMP-3, which has 477 residues including a 17-residue signal peptide. MMP-3 and collagenase are 54% identical in sequence, suggesting a common origin for the evolution of the two proteinases. MMP-3 and collagenase expression are coordinately modulated in synovial fibroblast cultures.1 MMP-3 is a secreted metalloprotease produced predominantly by connective tissue cells. Together with other metalloproteases, it can synergistically degrade the major components of the extracellular matrix. It is capable of degrading proteoglycan, fibronectin, laminin, and type IV collagen, but not interstitial type I collagen. MMP-3 genotype may be an important determinant of vascular remodeling and age-related arterial stiffening, with the heterozygote having the optimal balance between matrix accumulation and deposition.2 The standard product used in this kit is recombinant human MMP-3, consisting of 460 amino acids with the molecular mass of 52KDa. The detected MMP-3 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. Saus, J.; Quinones, S.; Otani, Y.; Nagase, H.; Harris, E. D., Jr.; Kurkinen, M. The complete primary structure of human matrix metalloproteinase-3: identity with stromelysin. J. Biol. Chem. 263: 6742-6745, 1988.
2. Medley, T. L.; Kingwell, B. A.; Gatzka, C. D.; Pillay, P.; Cole, T. J. Matrix metalloproteinase-3 genotype contributes to age-related aortic stiffening through modulation of gene and protein expression. Circ. Res. 92: 1254-1261, 2003.
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