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You are here:Home » Kits and Assays » Kits for Apoptosis, BioAssay™ » Calpain, Fluorometric BioAssay™ Kit

Calpain, Fluorometric BioAssay™ Kit

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Specifications

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 (lmax=400nM); upon cleavage of the substrate by calpain, free AFC emits a yellow-green fluorescence (lmax=505nM), 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.
Catalog #C1035-31B
Sample TypeCell and tissue lysates
Intended UseThe kit provides a simple and convenient means for analyzing calpain activity in apoptotic and other samples. The assay is based on fluorometric detection of the cleavage of calpain substrate at 505nm using a fluorometer or a fluorecence plate reader.
SpecificityMammalian
Absorption 400nm
Excitement 505nm
Fluorescence Ex/Em 400/505nm
Kit componentsExtraction Buffer 1x25ml
10X Reaction Buffer 1x1.5ml
Calpain Substrate Ac-LLY-AFC 1x500ul
Active Calpain I (Positive Control) 1x10ul
Calpain Inhibitor Z-LLY-FMK 1x10ul
Storage and StabilityStore liquids at -70°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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