Abcam, ab65308, Calpain Activity Assay Kit

Size: 100Test
Calpain Activity Assay Kit ab65308 provides optimized buffers and reagents for a convenient measurement of calpain activity. Readout on fluorometric plate reader. Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Key facts
Detection method:Fluorescent,
Sample types:Cell Lysate,
Assay type:Quantitative,
Assay time:2h,
Assay Platform:Microplate reader

Product details:
Calpain Activity Assay Kit ab65308 provides optimized buffers and reagents for a convenient measurement of calpain activity.
The extraction buffer included in the kit specifically extracts cytosolic proteins without contamination by cell membrane and lysosome proteases. It also prevents auto-activation of calpain during the extraction procedure. Thus, the kit detects only activated calpain within the cytosol.
The calpain activity assay protocol 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.
Calpain activity assay protocol summary
- add samples and positive and negative controls to wells
- add reaction buffer and calpain substrate
- incubate for 60 min
- analyze with a microplate reader
Other Notes
This product was previously called K240 Biovision Calpain Activity Fluorometric Assay Kit.
If additional Ac-LLY-AFC substrate is needed, it can be purchased separately as
ab171379
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions--80°C, Appropriate long-term storage conditions--80°C, Storage information-Please refer to protocols

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Calpain 1 also known as CAPN1 is a calcium-dependent cysteine protease with a mass of approximately 80 kDa. It exhibits calpain activity by cleaving substrate proteins in a regulated manner. Calpain 1 expresses in various tissues including muscle brain and lens suggesting its involvement in diverse cellular processes. The enzyme is part of a larger calpain family with calpain-1 calpain-2 and other isoforms that demonstrate differing substrate specificities and regulatory mechanisms.
Biological function summary
Calpain 1 participates in cellular functions such as signal transduction cell cycle progression and apoptosis. It is not part of a complex but operates independently to modulate cellular responses by processing different target proteins. Calpain protein activity impacts cytoskeletal remodeling and membrane repair important processes in cell mobility and stability. Through its proteolytic actions calpain 1 influences neural plasticity and muscle function demonstrating its broad biological significance.
Pathways
Calpain 1 functions within the calcium signaling and apoptotic pathways highlighting its role in regulating cellular survival and death. Calcium influx triggers calpain activity linking it to proteins such as calmodulin and calcineurin that also respond to calcium signals. Calpain-1 modulates apoptosis by targeting Bcl-2 family proteins influencing cell fate during stress responses. The calpain assay measures its activity providing insights into its contribution to these vital pathways.
Calpain 1 connects to neurodegenerative diseases like Alzheimer's and muscular diseases such as muscular dystrophy. Alzheimer's disease features abnormal calpain activity affecting tau protein and amyloid precursor protein processing. In muscular dystrophy calpain-1's unregulated activity leads to excessive muscle degradation aggravating the disease condition. Its interaction with spectrin and dystrophin proteins suggests a mechanistic role in the pathogenesis of these disorders pinpointing calpain-1 as a potential therapeutic target.