Calpain Activity Fluorometric Assay Kit

Activation of calpain is involved in many forms of physiological and pathological processes (e.g., apoptosis). Calpain activation requires cell membrane and Ca²ᶧ, 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 (λmax = 400 nm); upon cleavage of the substrate by calpain, free AFC emits a yellow-green fluorescence (λmax = 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.

• Detection method: Fluorescence (Ex/Em 400/505 nm) • Species reactivity: Mammalian • Applications: The 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 505 nm using a fluorometer or a fluorecence plate reader.

• Simple one-step procedure; takes only ~2 hours • Fast and convenient • The kit contains both positive and negative controls. Active calpain and calpain inhibitor and antibodies are also available separately.

Apoptosis Assay Kit

none

• Extraction Buffer • 10X Reaction Buffer • Calpain Substrate Ac-LLY-AFC • Active Calpain I (Positive Control) • Calpain Inhibitor Z-LLY-FMK

-70°C

dry ice

6-12 months

see datasheet

The 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 505 nm using a fluorometer or a fluorecence plate reader.

Cell and tissue lysates

Biovision supplies other types of Assays as 1.