| Solution Information | help | |
| Enzyme: | Collagenase 3 | |
| inhibitor: | BDBM40468 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: Forty-six compounds identified from a previously described set of experiments entitled "Primary biochemical high-throughput screening assay for inhibitors of Matrix Metalloproteinase 13 (MMP13) activity" (PubChem AID = 570) have been assessed in this dose response experiment. As with the primary screen, this assay is based on ability of MMP-13 to cleave a fluorogenic triple-helical peptide (fTHP), which has been modeled after the consensus binding and cleavage site recognized by MMP-13 in type I-III collagens. The fTHP is labeled with both a (7-methoxycoumarin-4-yl)acetyl fluorophore (Mca) and a 2,4-dinitrophenyl quencher (Dnp) that comprise a fluorescence energy transfer (FRET) pair. Cleavage of the fHTP peptide results in liberation of Mca fluorescence. In this assay, the fluorescence signal from unquenched Mca is directly proportional to MMP-13 activity. The dose-response assay was conducted in 1536 well plate format. Each compound was assayed in triplicate, | |
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