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Matrix metalloproteinase-3 (MMP-3) fluorometric drug discovery kit, GREEN

 
BML-AK303-0001 96 wells 500.00 USD
 

Product Specification

Application:Designed to screen MMP-3 inhibitors using a quenched fluorogenic substrate.
 
Handling:Avoid freeze/thaw cycles.
 
Long Term Storage:-80°C
 
Kit/Set Contains:1 vial MMP-3 enzyme
1 vial substrate (MMP-3 fluorogenic substrate)
1 vial 5-FAM calibration standard
1 vial control inhibitor (NNGH)
1 bottle (20 ml) assay buffer
1 black 96-well microplate
Instructions
 
Miscellaneous/General:Matrix metalloproteinase-3 (MMP-3, stromelysin-1, transin-1) is a member of the MMP family of extracellular proteases. These enzymes play a role in many normal and disease states by virtue of their broad substrate specificities. Targets of MMP-3 include collagens, fibronectin, and laminin, plasminogen, HB-EGF, E-cadherin, and other MMPs. MMP-3 is secreted as a 55-59kDa glycosylated proenzyme (measured by SDS-PAGE), and activated by cleavage to forms of 21-48kDa. It is unique from other MMPs in that its pH optimum is 5.9, rather than around 7.0.
 
The MMP-3 Fluorometric (also known as fluorimetric)Drug Discovery Kit, GREEN is a complete assay system designed to screen MMP-3 inhibitors using a quenched fluorogenic MMP-3 substrate: 5-FAM-Arg-Pro-Lys-Pro-Val-Glu-Nva-Trp-Arg-Lys(TQ2W)-NH2 [5-FAM=5-carboxyfluorescein; Nva=norvaline; TQ2W=quencher]. FAM fluorescence is thoroughly quenched by the TQ2W group until cleavage by MMPs separates the two moieties. 
 
The MMP-3 Fluorogenic Substrate offers key advantages over other MMP substrates. 1) Emission at the green end of the spectrum avoids the interference at lower wavelengths often exhibited by screening compounds, and by substances commonly found in biological samples and tissue culture medium. 2) The ultra-strong fluorescence of this substrate allow for substrate concentrations much lower than the Km, a condition generally desirable in inhibitor screening/kinetics assays.
 
The assays are performed in a convenient 96-well microplate format. The kit is useful to screen inhibitors of MMP-3, a potential therapeutic target. The compound NNGH is also included as a prototypic control inhibitor.
 
Background / Technical Information:NCBI Reference Sequence: NM_002422
The MMP-3 Fluorogenic Substrate offers key advantages over other MMP substrates. 1) Emission at the green end of the spectrum avoids the interference at lower wavelengths often exhibited by screening compounds, and by substances commonly found in biological samples and tissue culture medium. 2) The ultra-strong fluorescence of this substrate allow for substrate concentrations much lower than the Km, a condition generally desirable in inhibitor screening/kinetics assays.
 

General Literature References

Matrix metalloproteinases: regulators of the tumor microenvironment: K. Kessenbrock & Z. Werb; Cell 141, 52 (2010), Abstract;
Updated biological roles for matrix metalloproteinases and new "intracellular" substrates revealed by degradomics : G.S. Butler & C.M. Overall; Biochemistry 48, 10830 (2009), Abstract;
Matrix metalloproteinases: they're not just for matrix anymore!: L.J. McCawley & L.M. Matrisian; Curr. Opin. Cell Biol. 13, 534 (2001), Abstract;
Release of an invasion promoter E cadherin fragment by matrilysin and stromelysin: V. Noë, et al.; J. Cell Sci. 114, 111 (2001), Abstract;
A rationalization of the acidic pH dependence for stromelysin-1 (matrix metalloproteinase-3) catalysis and inhibition: L.L. Johnson, et al.; J. Biol. Chem. 275, 11026 (2000), Abstract; Full Text
Matrix metalloproteinase degradation of extracellular matrix: biological consequences: S.D. Shapiro; Curr. Opin. Cell Biol. 10, 602 (1998), Review, Abstract;
Discovery of CGS 27023A, a non-peptidic, potent, and orally active stromelysin inhibitor that blocks cartilage degradation in rabbits: L.J. MacPherson, et al.; J. Med. Chem. 40, 2525 (1997), Abstract;

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