Overview

Product name
Calpain Activity Assay Kit


Product overview
Activation of calpain is involved in many forms of physiological and pathological processes (e.g., apoptosis). Calpain activation requires cell membrane and Ca2+, and activated calpain is released into cytosol. The Calpain Activity Assay Kit provides optimized buffers and reagents for a convenient measurement of calpain activity. The Extraction Buffer provided with the kit specifically extracts cytosolic proteins without contaminations of cell membrane and lysosome proteases. The Extraction Buffer also prevents auto activation of calpain during the extraction procedure. Thus, the kit detects only activated calpain in cytosol upon treatment of cells with inducers (e.g., chemicals or drugs). The fluorometric assay is based on the detection of cleavage of calpain substrate Ac-LLY-AFC. Ac-LLY-AFC emits blue light (lambda max = 400 nm); upon cleavage of the substrate by calpain, free AFC emits a yellow-green fluorescence (lambda max = 505 nm), which can be quantified using a fluorometer or a fluorecence plate reader. Comparison of the fluorescence intensity from a treated sample with a normal control allows determination of the changes in calpain activity.


Notes
Store kit at –70oC (Store Extraction Buffer and 10X Reaction Buffer at 4oC after opening). All reagents are stable for 6 months under proper storage conditions.



Positive Control Active Calpain I is at a concentration of 1 ug/ul and has a specific activity of 1000 U/mg protein (unit definition: 

One unit is defined as the amount of enzyme that will hydrolyze one pmol Suc-LLVY-AMC per minute at 25 °C.)

Properties

Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.



    

        

            
Components
            
Identifier
            
100 tests
        
    
    

        

            
10X Reaction Buffer
            
Clear
            
1 x 1.5ml
        
        

            
Active Calpain I (Positive Control)
            
Green
            
1 x 10μl
        
        

            
Calpain Activity Assay Kit
            
ab65308
            
1 x 100 kits
        
        

            
Calpain Inhibitor Z-LLY-FMK
            
Orange
            
1 x 10μl
        
        

            
Calpain Substrate Ac-LLY-AFC
            
Amber
            
1 x 500μl
        
        

            
Extraction Buffer
            
WM
            
1 x 25ml
        
    


http://www.abcam.com/Calpain-Activity-Assay-Kit-ab65308.html