Calpain-Glo Protease Assay, Promega
Supplier: Promega Corporation
The Calpain-Glo Protease Assay is a homogeneous, luminescent assay that measures calpain 1 (µ) and 2 (m) activities
- Homogeneous, Luminescent Assay that Measures Calpain 1 (micro) and 2 (m) Activities
- Simple "add-mix-measure" protocol
- Maximum sensitivity while enzyme is still active
- 1,000x more sensitive than fluorometric assays
The Calpain-Glo Protease Assay is a homogeneous, luminescent assay that measures calpain 1 (u) and 2 (m) activities. Calpains are a family of calcium-activated cysteine proteases involved in cleaving a wide variety of proteins. Calpains modulate the biological activities of their substrates via limited proteolysis. The Calpain-Glo Protease Assay provides a succinyl, proluminescent calpain substrate, Suc-LLVY-aminoluciferin, in a buffer system optimized for calpain and luciferase activities. The addition of the calpain reagent in an add-mix-measure format results in calpain cleavage of the substrate and rapid development of a glow-type luminescent signal produced by the luciferase reaction. The signal is proportional to the amount of calpain activity present. The assay is designed for use with purified enzyme preparations.
It is up to 1000X more sensitive than traditional fluorometric assays because it avoids inherent fluorescent background signals. Maximum sensitivity can be reached in as little as 10 minutes, so the enzyme is still fully active. Results are linear over four logs of calpain concentration.
The assay provides a succinyl, proluminescent calpain substrate, Suc-LLVY-aminoluciferin, in a buffer system optimized for calpain and luciferin activities. The addition of the calpain reagent in an add-mix-measure format results in calpain cleavage of the substrate and rapid development of a luminescent signal produced by the luciferase reactions. The signal is proportional to the amount of calpain activity present. The assay is designed for use with purified enzyme preparations and can be used in a multiwell format.
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