posted 07-15-2011 08:04 PM            

The SensoLyte® 520MMP-15, MMP-16 and MMP-24 Assay Kits provide a convenient assay for high throughput screening (HTS) of inducers and inhibitors for MMP-15, MMP-16 and MMP-24, respectively. The substrate used in each of these kits detect MMP-15, 16 or 24 activity using a 5-FAM/QXL™ 520 FRET peptide. In the intact FRET peptide, the fluorescence of 5-FAM is quenched by QXL™ 520. Upon cleavage into two separate fragments by MMP-15, 16 or 24, the fluorescence of 5-FAM is recovered, and can be monitored at excitation/emission wavelengths = 490 nm/520 nm. With excellent fluorescence quantum yield and longer wavelength, 5-FAM shows less interference from auto fluorescence of test compounds and cellular components. The assays are performed in a convenient 96-well microplate format. The 5-FAM/QXL™ 520 substrates can detect subnanogram level of MMP-15, 16 and 24 activity.

AnaSpec is pleased to also release the SensoLyte® 390 Generic MMP Activity Assay Kit. This Kit is optimized to detect the activity of a variety of MMPs, including MMP-1, 2, 7, 8, 9, 13, 14, 15, 16, and 24. It can be used for detecting generic MMP activity in biological samples or for HTS of MMP inducers and inhibitors using purified MMPs. An Mca/Dnp FRET peptide is used as a substrate. In the intact FRET peptide, the fluorescence of Mca is quenched by Dnp. Upon cleavage into two separate fragments by MMPs, the fluorescence of Mca is recovered, and can be monitored at Ex/Em = 330/390 nm. The assays are also performed in a convenient 96-well microplate format.

The matrix metalloproteinases (MMPs) constitute a family of zinc-dependent endopeptidases that function within the extracellular matrix (ECM). These enzymes are responsible for the breakdown of connective tissues and are important in bone remodeling, the menstrual cycle, and repair of tissue damage.1, 2 MMP-15 (also known as MT2-MMP), MP-16 (MT3-MMP) and MMP-24 (MT5-MMP) are members of the membrane-type MMP (MT-MMP) subfamily. MMP-15 contains a potential transmembrane domain and is expressed at the cell surface rather than secreted.3 MMP-16 is found in soluble form, while MMP-24 may function as a soluble proteinase since it is shed from cell membrane.4 All these 3 MMPs activate pro-MMP-2 (gelatinase-A) that is involved in tumor development and metastasis, and thus are proposed as therapeutic targets for cancer.4-7

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References:
1. Visse, R. et al. Circ. Res. 92, 827 (2003).
2. Nagase, H. et al. Cardiovasc. Res. 69, 562 (2006).
3. Fillmore, HL. et al. J Neurooncol. 53, 187 (2001).
4. Amalinei, C. et al. Romanian J. Morph. Embry. 48, 323(2007).
5. Collier, IE. et al. J. Biol. Chem. 263, 6579 (1988).
6. Salo, T. et al. J. Biol. Chem. 258, 3058 (1983).
7. Salo, T. et al. J. Biol. Chem. 260, 8526 (1985).

AnaSpec, EGT Group

AnaSpec, EGT Group is a leading provider of integrated proteomics and genomics solutionsTM for worldwide life science research. AnaSpec offers expertise in peptides, detection reagents, antibodies, assay kits, amino acids, oligonucleotides, and qPCRs. AnaSpec carries a broad product line of biochemicals and reagents for basic research, high-throughput screening and drug discovery. AnaSpec also provides premier custom services including peptide synthesis, antibody production, assay development, fluorescent dyes, and oligonucleotide synthesis. AnaSpec is certified to ISO 9001:2008.

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