Overview
- 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.
Visit our FAQs page for tips and troubleshooting. - Notes
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.
- Tested applicationsFunctional Studies more details
Properties
- Storage instructionsStore at -20°C. Please refer to protocols.
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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
Contains 1 calpain catalytic domain.
Contains 4 EF-hand domains.
Target information above from: UniProt accession
P07384
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- Calcium activated neutral proteinase 1Calcium-activated neutral proteinase 1Calpain 1 (mu/I) large subunit
- Calpain 1 catalytic subunitCalpain 1 large subunitCalpain large polypeptide L1Calpain mu typeCalpain mu-typeCalpain-1 catalytic subunitCalpain-1 large subunitCAN1_HUMANCANPCANP 1CANP1CANPL1CAPN 1CAPN1Cell proliferation inducing gene 30 proteinCell proliferation inducing protein 30Cell proliferation-inducing gene 30 proteinMicromolar calpainMicromolar-calpainmuCANPmuCL
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Applications
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.
| Application | Notes |
|---|---|
| Functional Studies | FuncS |
Calpain Activity Assay Kit images
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Functional Studies - Calpain Activity Assay Kit (ab65308)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. -
Functional Studies - Calpain Activity Assay Kit (ab65308)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.
Protocols
References for Calpain Activity Assay Kit (ab65308)
This product has been referenced in:
- 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) »
- Das A et al. Post-treatment with voltage-gated Na(+) channel blocker attenuates kainic acid-induced apoptosis in rat primary hippocampal neurons. Neurochem Res 35:2175-83 (2010). Functional Studies ; Rat . Read more (PubMed: 21127971) »
