Calpain Activity Assay Kit ab65308 provides optimized buffers and reagents for a convenient measurement of calpain activity.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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- Arthritis research & therapy 25:2122023ACSL4 inhibition prevents macrophage ferroptosis and alleviates fibrosis in bleomycin-induced systemic sclerosis model.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDianyu Cao,Jina Zheng,Zheng Li,Yong Yu,Zengrui Chen,Qiang WangPubMed 37884942
- Arthritis research & therapy 25:1172023Aberrant mechanical loading induces annulus fibrosus cells apoptosis in intervertebral disc degeneration via mechanosensitive ion channel Piezo1.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChenhao Liu,Xiaoxin Gao,Jinhui Lou,Haiyin Li,Yuxuan Chen,Molong Chen,Yuyao Zhang,Zhilei Hu,Xian Chang,Menglin Luo,Yu Zhai,Changqing LiPubMed 37420255
- Heliyon 9:e156112023Changes in beef protein digestibility in an infant digestion model with prefreezing temperatures and aging periods.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSeonmin Lee,Kyung Jo,Hyun Gyung Jeong,Yun-Sang Choi,Samooel JungPubMed 37153398
- Cancer cell international 23:492023Changes in calpain-2 expression during glioblastoma progression predisposes tumor cells to temozolomide resistance by minimizing DNA damage and p53-dependent apoptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMaren Nicole Stillger,Chia-Yi Chen,Zon Weng Lai,Mujia Li,Agnes Schäfer,Axel Pagenstecher,Christopher Nimsky,Jörg Walter Bartsch,Oliver SchillingPubMed 36932402
- Molecular neurobiology 60:3311-33282023Extracellular CIRP Induces Calpain Activation in Neurons via PLC-IP-Dependent Calcium Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesArchna Sharma,Ezgi Sari,Yongchan Lee,Shivani Patel,Max Brenner,Philippe Marambaud,Ping WangPubMed 36853429
- Journal of cellular and molecular medicine 27:232-2452022Experimental diabetes exacerbates autophagic flux impairment during myocardial I/R injury through calpain-mediated cleavage of Atg5/LAMP2.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLichun Guan,Ziqin Yu,Zhimei Che,Hang Zhang,Yong Yu,Dicheng Yang,Dewei Qian,Ruizhen Chen,Min YuPubMed 36562207
- Cancer medicine 12:8378-83872022Mistletoe lectin inhibits growth of Myc-amplified small-cell lung cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMohammad A Shatat,Betsy Gauthier,Suzy Yoon,Eric Yuan,Peiying Yang,Goutham Narla,Afshin Dowlati,Richard T LeePubMed 36562288
- International journal of molecular sciences 23:2022Distinct Changes in Calpain and Calpastatin during PNS Myelination and Demyelination in Rodent Models.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJohn A Miller,Domenica E Drouet,Leonid M Yermakov,Mahmoud S Elbasiouny,Fatima Z Bensabeur,Michael Bottomley,Keiichiro SusukiPubMed 36499770
- Neuron 111:30-48.e142022Mechanosensitive brain tumor cells construct blood-tumor barrier to mask chemosensitivity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXin Chen,Ali Momin,Siyi Wanggou,Xian Wang,Hyun-Kee Min,Wenkun Dou,Zheyuan Gong,Jade Chan,Weifan Dong,Jerry J Fan,Yi Xiong,Kamilia Talipova,Hongyu Zhao,Yuki X Chen,Kelly Veerasammy,Adam Fekete,Sachin A Kumar,Hongwei Liu,Qi Yang,Joe Eun Son,Zhengchao Dou,Malini Hu,Parnian Pardis,Kyle Juraschka,Laura K Donovan,Jiao Zhang,Vijay Ramaswamy,Hayden J Selvadurai,Peter B Dirks,Michael D Taylor,Lu-Yang Wang,Chi-Chung Hui,Rinat Abzalimov,Ye He,Yu Sun,Xuejun Li,Xi HuangPubMed 36323321
- Arthritis research & therapy 24:1482022Myeloid cell-specific deletion of Capns1 prevents macrophage polarization toward the M1 phenotype and reduces interstitial lung disease in the bleomycin model of systemic sclerosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLi Zhang,Dong Zheng,Yuemei Yan,Yong Yu,Ruizhen Chen,Zheng Li,Peter A Greer,Tianqing Peng,Qiang WangPubMed 35729674
Key facts
- Detection method
Fluorescent
- Sample types
Cell Lysate
- Assay type
Quantitative
- Reactive species
Mammals
- Assay time
2h
Target data
Function
Calcium-regulated non-lysosomal thiol-protease which catalyzes limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction (PubMed:21531719, PubMed:2400579). Proteolytically cleaves CTBP1 at 'Asn-375', 'Gly-387' and 'His-409' (PubMed:23707407).
Alternative names
Calpain-1 catalytic subunit, Calcium-activated neutral proteinase 1, Calpain mu-type, Calpain-1 large subunit, Cell proliferation-inducing gene 30 protein, Micromolar-calpain, CANP 1, muCANP, CAPN1, PIG30, CANPL1
What's included?
Recommended products
Calpain Activity Assay Kit ab65308 provides optimized buffers and reagents for a convenient measurement of calpain activity.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Alternative names
Calpain-1 catalytic subunit, Calcium-activated neutral proteinase 1, Calpain mu-type, Calpain-1 large subunit, Cell proliferation-inducing gene 30 protein, Micromolar-calpain, CANP 1, muCANP, CAPN1, PIG30, CANPL1
Key facts
- Detection method
Fluorescent
- Sample types
Cell Lysate
- Assay type
Quantitative
- Reactive species
Mammals
- Assay time
2h
- Assay Platform
Microplate reader
Storage
- 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
Notes
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
This product is manufactured by BioVision, an Abcam company and was previously called K240 Calpain Activity Fluorometric Assay Kit. K240-100 is the same size as the 100 test size of ab65308.
If additional Ac-LLY-AFC substrate is needed, it can be purchased separately as ab171379.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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3 product images
Functional Studies - Calpain Activity Assay Kit (ab65308)
Different amounts of positive control (Calpain I) treated with 1 μL of inhibitor (Z-LLY-FMK), background signal subtracted, duplicates; +/- SD.
Functional Studies - Calpain Activity Assay Kit (ab65308)
10e7 Jurkat cells (in 10 mL) were cultured in the absence or presence of 10 μM Camptothecin (CPT) (Camptothecin, DNA topoisomerase inhibitor ab120115) or 10 μg/mL Cycloheximide (CHX) (Cycloheximide, Protein synthesis inhibitor ab120093) for 4 hours. Pelleted cells were lysed in 0.5 mL of Extraction Buffer and tested directly. Background signal subtracted, duplicates; +/- SD.
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.
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