Product Overview :
The MMP-10 Fluorometric Drug Discovery Kit is a complete assay system designed to screen MMP-10 inhibitors using a quenched fluorogenic peptide: fluorogenic substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 [Mca=(7- methoxycoumarin-4-yl)-acetyl; Dpa=N-3-(2,4-dinitrophenyl)-L- α-β-diaminopropionyl]. Mca fluorescence is quenched by the Dpa group until cleavage by MMPs at the Gly-Leu bond separates the two moieties. The assays are performed in a convenient 96-well microplate format. The kit is useful to screen inhibitors of MMP-10, a potential therapeutic target. The compound NNGH11 is also included as a prototypic control inhibitor.
Kit Components :
MMP-10 ENZYME (HUMAN, RECOMBINANT)
FORM: E. coli recombinant human MMP-10 catalytic domain (calculated MW 19.4 kDa), 5 U/µl.
UNIT DEFINITION: One unit is defined as the amount of enzyme that will hydrolyze 100uM thiopeptide Ac-PLG-[2-mercapto-4-methyl-pentanoyl]-LG-OC2H5 at 100 pmol/min at 37°C.
STORAGE: -70°C; Avoid freeze/thaw cycles
QUANTITY: 150 U
PRESENTATION: 30 µl in screw-cap microfuge vial.
SUBSTRATE (fluorogenic substrate peptide; MW=1093.2)
FORM: 400 uM (437 ug/ml) in DMSO (dimethylsulfoxide)
STORAGE: -70°C
PRESENTATION: 200 ul in amber screw-cap microfuge vial.
CALIBRATION STANDARD (fluorogenic control peptide, MCA-Pro-Leu-OH; MW=444.5)
FORM: 40 uM (17.8 ug/ml) in DMSO
STORAGE: -70°C
PRESENTATION: 50 ul in amber screw-cap microfuge vial.
INHIBITOR (NNGH; MW=316.4)
FORM: 1.3mM in DMSO
STORAGE: -20 or -70°C
PRESENTATION: 50ul in screw-cap microfuge vial.
ASSAY BUFFER
50mM HEPES, 10mM CaCl2, 0.05% Brij-35, pH 7.5
FORM: Liquid in screw-cap plastic bottle
STORAGE: Room temperature
QUANTITY: 20 ml
½ VOLUME 96-WELL WHITE NBS MICROPLATE
STORAGE: Room temperature.
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