EnSens® Fluorescent 3D Live Cell Imaging Kit for MMP-3/MMP-10 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-3/MMP-10 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-3/MMP-10 Activity Detection Substrate: Despite the highly-overlapping substrate specificities of MMPs, EnSens® substrates are designed for enhanced selectivity. The graph below shows that at pH 6.0, the EnSens® MMP-3/MMP-10 activity detection substrate is resistant to most MMPs, although it is cleaved by MMP-2, MMP-9, and MMP-13 in addition to MMP-3 and MMP-10 (light blue bars). Simple addition of the commercially available inhibitor SB-3CT to the assay makes it selective for MMP-3 and MMP-10 (dark blue bars).
•EnSens® MMP-3/MMP-10 Activity Detection Substrate
•Far Red Fluorogen (FRF dye)
•Pre-activated substrate (EnSens®-PA, substrate positive control)
MMP-3 and MMP-10 are both known as stromelysins and possess significant overlap in their substrate recognition profiles. (MMP-11 is the third stromelysin and recognizes a largely unique set of substrates.) MMP-3 and MMP-10 are involved in diverse functions such as wound and muscle repair, infection, cancer, stroke, and arthritis. In addition to degrading extracellular matrix and basement membrane proteins such as gelatin, laminin, aggrecan, and fibronectin, they can also activate pro-MMPs. Other known targets for MMP-3 are decorin, IL-1beta, and pro-TNF-alpha.
EnSens Fluorescent 3D Live Cell Imaging Kit for MMP-3/MMP-10 Activity Tracking
- Product Code: 022315_10
- Availability: In Stock