EnSens® MMP-13 Activity Detection Kit Description
Pricing (select from Available Options, above):
Trial size (25 assays) $169
96 assays $675
384 assays $1039
Also available through VWR
Mix-and-Read, Fluorescence based Protease Activity detection kit. The kit includes protease selective detection reagent, fluorescent dye, reaction buffer, and substrate positive control. Protocol volume can be easily scaled up or down as needed. It is designed for fluorimetric endpoint and kinetic microplate assays. Quantifiable down to nano- and picomolar range of active protease concentrations. Upon activation, se-Red reagent produces detectable fluorescent signal in the far red region of the light spectrum, reducing background signal and increasing dynamic range. Excitation 625-635nm/ Emission 655-665nm.
Selectivity of the EnSens® MMP-13 Activity Detection Substrate: Despite the highly-overlapping substrate specificities of the collagenase MMPs (MMP-1, MMP-8, and MMP-13), the EnSens® MMP-13 substrate is cleaved by MMP-13, but not by MMP-1 or MMP-8. The substrate is cleaved somewhat by MMP-2, -3, -10, and 12 in addition to MMP-13, but the MMP-13-selective (and commercially available) inhibitor CL-82198 can be included in the assay to optimize the selectivity of the substrate.
The graph on the left shows selective inhibition of MMP-13 by CL-82198 when 50nM of each enzyme was assayed separately.
In the graph on the right, mixtures of MMPs (4nM each of MMP-2, MMP-3, MMP-10, MMP-12, with or without 4nM MMP-13) were assayed in the absence or presence of CL-82198. The inhibitor makes little to no difference in total activity when MMP-13 is not included in the mixture (light-colored bars). When MMP-13 is included in the mixture, CL-82198 significantly reduces the total activity (dark-colored bars).
This demonstrates that, with the use of MMP-13 inhibitor CL-82198, the EnSens MMP-13 substrate can selectively measure MMP-13 enzyme activity in a mixture of MMPs, either qualitatively or quantitatively (using a standard curve built from purified enzyme). For reference to this inhibitor strategy, see Berger AB et al. 2006 Identification of early intermediates of caspase activation using selective inhibitors and activity-based probes. Mol Cell 23: 509-521; and Corasaniti MT et al. 2007 Cell signaling pathways in the mechanisms of neuroprotection afforded by bergamot essential oil against NMDA-induced cell death in vitro. Br J Pharmacol. 151: 518-29.
•EnSens® MMP-13 Activity Detection Substrate
•Far Red Fluorogen (FRF dye)
•Pre-activated substrate (EnSens®-PA, substrate positive control)
•2X Assay buffer
•Molecular biology grade water
MMP-13 is one of the three collagenase matrix metalloproteases (MMP-1, MMP-8, and MMP-13). By virtue of its substrate profile (extracellular matrix and connective tissue proteins such as collagen, gelatin, aggrecan, decorin, and fibronectin), it is involved in arthritis and cancer/metastasis. It also cleaves and activates TGF-beta.
EnSens MMP-13 Activity Detection Kit
- Product Code: 032014_11
- Availability: In Stock