MMP-1 (interstitial collagenase, fibroblast collagenase) is an extracellular protease. This enzyme plays a significant role in cancer, arthritis and other diseases, and has been identified as a drug-screening target. SensoLyte® 520 MMP-1 Assay Kit uses a 5-FAM (fluorophore) and QXL520™ (quencher) labeled FRET peptide substrate for continuous measurement of enzyme activity. In an intact FRET peptide, the fluorescence of 5-FAM is quenched by QXL520™. Upon the cleavage of the FRET peptide by MMP-1, 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/QXL520™ based FRET peptide has less interference from the autofluorescence of test compounds and cellular components and provides better assay sensitivity. The assays are performed in a convenient 96-well or 384-well microplate format. 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-1 activity, please choose the SensoLyte® Plus MMP-1 Assay Kit, Cat# AS-72012.
Component A: MMP-1 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.