Matrix metalloproteinases (MMPs) belong to a family of secreted or membrane-associated proteins capable of digesting extracellular matrix components. MMP-13 (collagenase-3, rat collagenase) is involved in quite a few diseases such as arthritis and asthma. SensoLyte® 520 MMP-13 Assay Kit uses a 5-FAM (fluorophore) and QXL™ 520 (quencher) labeled FRET peptide substrates for continuous measurement of the enzyme activities. In an intact FRET peptide, the fluorescence of 5-FAM is quenched by QXL™ 520. Upon the cleavage of the FRET peptide by MMP-13, the fluorescence of 5-FAM is recovered, and can be continuously monitored at excitation/emission = 490 nm/520 nm. With superior fluorescence quantum yield and longer emission wavelength, 5-FAM/QXL™ 520 based FRET peptide is less interfered by the autofluorescence of test compounds and cellular components and provides better assay sensitivity. Members of the MMP family have poor substrate sequence specificity, making it difficult to use a peptide substrate alone to differentiate the activity of a particular MMP from other MMPs. If several MMPs are coexisting in your samples and you would like to specifically measure MMP-13 activity, please choose the SensoLyte® Plus MMP-13 Assay Kit, Cat# AS-72019.
Component A: MMP-13 substrate Ex/Em=490 nm/520 nm upon cleavage: 60 µL
Component B: 5-FAM-Pro-Leu-OH, Fluorescence reference standard Ex/Em=490 nm/520 nm: 1 mM, 10 µL
Component C: APMA, 4-aminophenylmercuric acetate (Caution: Organic mercury. Handle with care! Do not dispose into sink!): 1 M, 20 µL
Component D: Assay buffer: 20 mL
Component E: Stop solution: 10 mL
Storage & stability
Store all components at -20°C. Protect Components A and B from light. Components D and E can be stored at 4°C for convenience.