EnSens® Fluorescent 3D Live Cell Imaging Kit for Thrombin 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 Thrombin 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® Thrombin Activity Detection Substrate: Despite the highly-overlapping substrate specificities of Thrombin and Factor Xa, EnSens® substrates are able to differentiate between the two enzymes. In the graph below, the EnSens® Thrombin activity detection substrate is cleaved extremely well by Thrombin, but not at all by Factor Xa.
•EnSens® Thrombin Activity Detection Substrate
•Far Red Fluorogen (FRF dye)
•Pre-activated substrate (EnSens®-PA, substrate positive control)
(Coagulation factor IIa, fibrinogenase)
A clotting cascade protease initially expressed as inactive prothrombin and activated by factor Xa, thrombin cleaves fibrinogen at Arg-Gly, generating insoluble fibrin and fibrinopeptides A and B. It also cleaves and activates other clotting factor proteases, as well as clotting cascade inhibitor protein C. Thrombin is inactivated by the serpin antithrombin, and it is the target of the drug dabigatran (Pradaxa/Pradax/Prazaxa).
Together, thrombin and factor Xa proteases play prominent roles in the coagulation cascade and are major targets for emerging anti-coagulant therapies and associated diagnostics. EnSens fluorimetric reagents enable selective tracking and quantification of Factor Xa and Thrombin activation in purified protease, plasma, and whole blood samples.
EnSens Fluorescent 3D Live Cell Imaging Kit for Thrombin Activity Tracking
- Product Code: 022315_06
- Availability: In Stock