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

Calpain, Fluorometric BioAssay™ Kit


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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
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.
Sample TypeCell and tissue lysates
Absorption 400nm
Excitement 505nm
Fluorescence Ex/Em 400/505nm
Kit Components*C1035-31B1: Extraction Buffer, 1x25ml
*C1035-31B2: Reaction Buffer 10X, 1x1.5ml
C1035-31B3: Calpain Substrate Ac-LLY-AFC, 1x500ul
C1035-31B4: Active Calpain I (Positive Control), 1x10ul
C1035-31B5: Calpain Inhibitor Z-LLY-FMK, 1x10ul
Storage and StabilityStore components at -70°C. Once *C1035-31B1 and *C1035-31B2 are opened, store at 4°C. Stable for 6 months after receipt. 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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