EnSens® Fluorescent 3D Live Cell Imaging Kit for MMP-25 Activity Tracking
Pricing (select from Available Options, above):
Trial size (25 assays) $159
96 assays $599
The EnSens® Fluorescent 3D Live Cell Imaging Kit for MMP-25 Activity Tracking is a fluorescence based protease activity detection kit for use with live three-dimensional (3D) cell culture or co-culture. The kit provides non-toxic, protease-selective substrate and far-red fluorescent dye to be mixed directly into invasion matrix / scaffold / hydrogel (e.g. Matrigel®, or collagen gel), enabling real-time monitoring of specific protease activity by fluorescent microscopy. Kit components are stable for at least six days in culture, and highly resistant to photobleaching. Custom substrate development also available. Excitation 625-635nm/ Emission 655-665nm. Cy5 or similar filters can also be used.
Imaging the activity of a specific protease in live cells: The micrographs below demonstrate fluorescent live-cell 3D imaging of protease activity (in this case using the EnSens® MMP-14 Activity Detection Substrate) in an invading MDA-MB-231/fibroblast spheroid. Spheroids were formed in the wells of a Corning® 96-well Spheroid Microplate, then surrounded with a layer of Matrigel® Matrix. Data were generated by BioTek® Instruments using their CytationTM 5 Cell Imaging Multi-mode Reader (brightfield and Cy5 channels) and Gen5TM data analysis software.
Selectivity of the EnSens® MMP-25 Activity Detection Substrate: The EnSens® MMP-25 activity detection substrate demonstrates how EnSens® technology can generate new, selective protease substrates. Until now there were no known MMP-25 substrates, but the EnSens® MMP-25 substrate is cleaved extremely well by MMP-25 (kcat/Km 1.25 x 105 M-1s-1). It is also cleaved quite well by MMP-17 (MT4-MMP), likely because they are extremely closely related enzymes, being the only GPI (glycosyl-phosphatidylinositol)-anchored membrane-type MMPs, with very similar substrate specificities. However, note the selectivity of the MMP-25 substrate against the remaining MT-MMPs (MMP-14/MT1-MMP, MMP-15/MT2-MMP, MMP-16/MT3-MMP, MMP-24/MT5-MMP).
•EnSens® MMP-25 Activity Detection Substrate
•Far Red Fluorogen (FRF dye)
•Pre-activated substrate (EnSens®-PA, substrate positive control)
(MMP-25, Membrane-Type 6 Matrix Metalloproteinase, MT6-MMP, MT-MMP6, matrix metalloprotease-25, leukolysin)
A membrane-anchored metalloproteinase that can also be released as a soluble active form, MMP-25 is involved in diverse functions such as periodontal disease, arthritis, angiogenesis, tumor invasion and metastasis, and asthma. In addition to degrading extracellular matrix proteins such as collagen, fibronectin, and vimentin, it can also act on non-matrix proteins such as alpha 1 antitrypsin (mediating neutrophil migration) and chemokines. It also cleaves and activates its family member, MMP-2, further enhancing tumor invasion.
EnSens Fluorescent 3D Live Cell Imaging Kit for MMP-25 Activity Tracking
- Product Code: 022315_04
- Availability: In Stock