Calpain Activity Assay试剂盒(ab65308)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 2 hr
- Sample type: Cell Lysate, Tissue Extracts
概述
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产品名称
Calpain Activity Assay试剂盒 -
检测方法
Fluorescent -
样品类型
Tissue Extracts, Cell Lysate -
检测类型
Quantitative -
检测时间
2h 00m -
种属反应性
与反应: Mammals, Other species -
产品概述
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 -
说明
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.
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平台
Microplate reader
性能
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存放说明
Store at -80°C. Please refer to protocols. -
组件 100 tests 10X Reaction Buffer 1 x 1.5ml Calpain I Positive Control 1 x 10µl Calpain Inhibitor 1 x 10µl Calpain Substrate 1 x 500µl Extraction Buffer III 1 x 25ml -
研究领域
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功能
Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction. -
组织特异性
Ubiquitous. -
序列相似性
Belongs to the peptidase C2 family.
Contains 1 calpain catalytic domain.
Contains 4 EF-hand domains. -
细胞定位
Cytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding. - Information by UniProt
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别名
- Ca2 activated neutral protease
- Calcium activated neutral proteinase
- Calcium activated neutral proteinase small subunit
see all
图片
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Different amounts of positive control (Calpain I) treated with 1 μL of inhibitor (Z-LLY-FMK), background signal subtracted, duplicates; +/- SD.
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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.
数据表及文件
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SDS download
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Datasheet download
文献 (74)
ab65308 被引用在 74 文献中.
- Chen X et al. Mechanosensitive brain tumor cells construct blood-tumor barrier to mask chemosensitivity. Neuron 111:30-48.e14 (2023). PubMed: 36323321
- Miller JA et al. Distinct Changes in Calpain and Calpastatin during PNS Myelination and Demyelination in Rodent Models. Int J Mol Sci 23:N/A (2022). PubMed: 36499770
- Wu Y et al. Calpain Inhibitor Calpeptin Alleviates Ischemia/Reperfusion-Induced Acute Kidney Injury via Suppressing AIM2 Inflammasome and Upregulating Klotho Protein. Front Med (Lausanne) 9:811980 (2022). PubMed: 35155498
- Tao Y et al. Enhanced Orai1-mediated store-operated Ca2+ channel/calpain signaling contributes to high glucose-induced podocyte injury. J Biol Chem 298:101990 (2022). PubMed: 35490782
- Yang HH et al. Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs. Int J Mol Sci 23:N/A (2022). PubMed: 35163312