资料参考
Apoptosis is one of the biological processes that activation of calpain is implicated in. Cell membrane and Ca2+is required for calpainactivation. The activated calpain is entered into cytosol. The Calpain Activity Assay Kit is a convenient way of measuring calpain activity with optimized buffers and reagents. The Extraction Buffer extracts cytosolic proteins with contaminations from cell membrane and lysosome proteases. It also protects the cell from atuo-activation of calpain during extraction process. Thus, the kit only detects activated calpain in cytosol following treatment of cells with chemicals or drugs inducers. 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 fluorescence.
Features & Properties
Features | 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. |
Shipping | Dry ice |
Storage Conditions | Store at -70°C. |
概述
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. |
Kit components | Extraction Buffer
10X Reaction Buffer
Calpain Substrate Ac-LLY-AFC
Active Calpain I (Positive Control)
Calpain Inhibitor Z-LLY-FMK |