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Matrix metalloproteinase-12 (MMP-12) fluorometric drug discovery kit, GREEN

 
BML-AK312-0001 96 wells 525.00 USD
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The MMP-12 Fluorometric (also known as fluorimetric) Drug Discovery Kit, GREEN is a complete assay system designed to screen MMP-12 inhibitors using a quenched fluorogenic OMNIMMP® Green  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 assays are performed in a convenient 96-well microplate format. The kit is useful to screen inhibitors of MMP-12, a potential therapeutic target. The compound NNGH is also included as a prototypic control inhibitor.

Product Specification

Alternative Name:Metalloelastase, Macrophage elastase
 
Applications:Fluorescent detection, HTS
Activity assay
 
Application Notes:Designed to screen MMP-12 inhibitors using a quenched fluorogenic substrate.
 
Handling:Avoid freeze/thaw cycles.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Kit/Set Contains:1 vial MMP-12 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
 
Scientific Background:Matrix metalloproteinase-12 (MMP-12, metalloelastase, macrophage elastase, commonly confused with neutrophil elastase) 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-12 include elastin, fibronectin, laminin, plasminogen, u-PAR, and tissue factor pathway inhibitor. MMP-12 is secreted as a 53kDa proenzyme (as measured by SDS-PAGE), and activated by cleavage to forms of 45-22kDa.  MMP-12 is an important target for inhibitor screening due to its involvement in diseases such as cancer, allergy and emphysema.
 
Technical Info/Product Notes:NCBI Reference Sequence: NM_004994
The green fluorogenic MMP substrate provided in the kit offers key advantages over other MMP substrates. 1) Emission in the green region 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 allows for substrate concentrations much lower than the Km, a condition generally desirable in inhibitor screening/kinetics assays.
 
UniProt ID:P39900
 

General Literature References

Matrix metalloproteinase-12 is a therapeutic target for asthma in children and young adults: S. Mukhopadhyay et al.; J. Allergy Clin. Immunol. 126, 70 (2010), Abstract;
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;
Proteolysis of the urokinase-type plasminogen activator receptor by metalloproteinase-12: implication for angiogenesis in fibrin matrices: P. Koolwijk et al.; Blood 97, 3123 (2001), Abstract;
Matrix metalloproteinases cleave tissue pathway inhibitor. Effects on coagulation: A. Belaaouaj et al.; J. Biol .Chem. 275, 27123 (2000), Abstract;
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;
Hydrolysis of a broad spectrum of extracellular matrix proteins by human macrophage elastase: T.J. Gronski, Jr. et al.; J. Biol. Chem 272, 12189 (1997), Abstract;

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