Calpain Activity Assay Kit (ab65308)
- Product nameCalpain Activity Assay Kit
- Tests100 x 1 assay
- Sample typeTissue Extracts, Cell Lysate
- Assay typeQuantitative
- Assay time2h 00m
- Product overview
Abcam's 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.
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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.
- Tested applicationsFunctional Studies more details
- Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests 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
- Research Areas
- FunctionCalcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
- Tissue specificityUbiquitous.
- Sequence similaritiesBelongs to the peptidase C2 family.
Contains 1 calpain catalytic domain.
Contains 4 EF-hand domains.
- Cellular localizationCytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding.
- Calcium activated neutral proteinase 1
- Calcium-activated neutral proteinase 1
- Calpain 1 (mu/I) large subunit
- Calpain 1 catalytic subunit
- Calpain 1 large subunit
- Calpain large polypeptide L1
- Calpain mu type
- Calpain mu-type
- Calpain-1 catalytic subunit
- Calpain-1 large subunit
- CANP 1
- CAPN 1
- Cell proliferation inducing gene 30 protein
- Cell proliferation inducing protein 30
- Cell proliferation-inducing gene 30 protein
- Micromolar calpain
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.
Calpain Activity Assay Kit images
Typical Data for ab65308: Active Calpain (1 µg) was incubated at 37 °C for one hour using the Calpain Substrate with or without 20 µM Calpain Inhibitor.
References for Calpain Activity Assay Kit (ab65308)
This product has been referenced in:
- Mattheij NJ et al. Dual mechanism of integrin aIIbß3 closure in procoagulant platelets. J Biol Chem 288:13325-36 (2013). Read more (PubMed: 23519467) »
- Liu D et al. Activation of calpains mediates early lung neutrophilic inflammation in ventilator-induced lung injury. Am J Physiol Lung Cell Mol Physiol 302:L370-9 (2012). Functional Studies ; Mouse . Read more (PubMed: 22140070) »