Tissue-type plasminogen activator (tPA) is a serine protease that cleaves pro-enzyme plasminogen into active plasmin. It has been recognized that tPA play important roles in various physiological and pathological processes. In plasma, it functions as a thrombolytic enzyme and can be used for treatment of ischemic stroke. In the central nervous system, tPA promotes events associated with synaptic plasticity and cell death in a number of settings, such as cerebral ischemia and seizures. The SensoLyte AMC tPA Activity Assay Kit is optimized for assaying tPA enzyme activities. This kit contains a fluorogenic substrate with a high reactivity and low background. tPA cleaves the substrate resulting in release of AMC (7-amino-4-methylcoumarin) fluorophore. Fluorescence can be monitored at excitation /emission= 354/442 nm. This kit can be used for high throughput screening of tPA inducers and inhibitors.
Component A: tPA substrate, Ex/Em=354/442 nm upon substrate cleavage: 4 mM, 50 µL
Component B: AMC, fluorescence reference standard, Ex/Em=354/442 nm: 4 mM, 10 µL
Component C: 2X Assay Buffer: 10 mL
Component D: Purified tPA enzyme: 25 Units/µL, 40 µL
Component E: tPA Inhibitor: 10 mM, 100 µL
Storage & stability
Store all kit components, except Component D, at -20°C. Store Component D at -80°C. Aliquot as needed to avoid multiple freeze-thaw cycles. Protect Components A and B from light and moisture. Components C can be stored at room temperature for convenience.