Calpain-Glo Protease Assay

Description

• Overview
• Protocols
• Specifications
• Resources

• Simple Protocol Offers Fast, Sensitive Results

  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.

  • Designed for measuring calpain activity and/or inhibition in a multiwell format and easy to automate.
  • Well suited for rapidly autolysed enzymes like calpain; maximum sensitivity is reached in as little as 10 minutes, while the enzyme is fully active.
  • Up to 1,000 times more sensitive than competitive fluorometric assays. The luminescent assay avoids inherent fluorescent background signals, providing excellent signal-to-background readings. The assay is linear over 4 logs of calpain concentration.

  Reference
  

  1. Moldoveanu, T. et al. (2002) Cell 108, 649–60.

  

• Protocols

  Complete Protocol

  

  Calpain-Glo™ Protease Assay Technical Bulletin

  PDF (404 KB)

• Certificate of Analysis

  Lookup Certificate of Analysis

  Storage Conditions

  -30C TO -10C

  Use Restrictions

  For Research Use Only. Not for Use in Diagnostic Procedures.

• Resources

  

  Articles

  • Screen for Calpain Inhibitors Using a Cell-Based High-Throughput Assay
  • A Bioluminescent Assay for Calpain Activity
  

  Citations

  • Bioluminescent assays for ADMET
    --
    2008 Expert Opin. Drug Metab. Toxicol.