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Abcam's Calpain Activity Assay试剂盒provides optimized缓冲器and试剂for a convenient measurement of calpain activity. The Extraction缓冲器provided with the试剂盒specifically extracts cytosolic proteins without contaminations of cell膜and lysosome proteases. The Extraction缓冲器also prevents auto-activation of calpain during the extraction procedure. Thus， the试剂盒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.
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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.
If additional Ac-LLY-AFC substrate is needed, it can be purchased separately as ab171379.
|10X Reaction Buffer||Clear||1 x 1.5ml|
|Active Calpain I (Positive Control)||Green||1 x 10µl|
|Calpain Inhibitor Z-LLY-FMK||Orange||1 x 10µl|
|Calpain Substrate Ac-LLY-AFC||Amber||1 x 500µl|
|Extraction Buffer||WM||1 x 25ml|
Our Abpromise guarantee covers the use of ab65308 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent concentration.|
Different amounts of positive control (Calpain I) treated with 1 μL of inhibitor (Z-LLY-FMK), background signal subtracted, duplicates; +/- SD.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"