Overview

  • Product nameCaspase 3 Assay Kit (Fluorometric)See all Caspase 3 kits ...
  • Tests
    100 x 1 assay
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeEnzyme activity
  • Assay time
    2h 00m
  • Product overview

    Abcam's Caspase 3 Assay Kit (Fluorometric) provides a simple and convenient means for assaying the DEVD-dependent caspase activity. The assay is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). DEVD-AFC emits blue light (?max = 400 nm); upon cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (?max = 505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3/CPP32 activity.
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  • Notes

    Activation of ICE-family proteases/caspases initiates apoptosis or other cellular processes in mammalian cells.

  • Tested applicationsFunctional Studiesmore details

Properties

  • RelevanceThe caspase family of cysteine proteases play a key role in apoptosis. Caspase 3 (also known as CPP32, YAMA and apopain) is the most extensively studied apoptotic protein among caspase family members. Caspase 3 is synthesized as an inactive pro enzyme that is processed in cells undergoing apoptosis by self proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10). The processed form of Caspase 3 consists of large (17kD) and small (12kD) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28 - Ser29 and Asp175 - Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Alternative splicing of this gene results in two transcript variants which encode the same protein. In immunohistochemical studies Caspase 3 expression has been shown to be widespread but not present in all cell types (e.g. commonly reported in epithelial cells of skin, renal proximal tubules and collecting ducts). Differences in the level of Caspase 3 have been reported in cells of short lived nature (eg germinal centre B cells) and those that are long lived (eg mantle zone B cells). Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease.
  • Cellular localizationCytoplasmic
  • Alternative names
    • Apopain
    • Apopain precursor
    • CASP 3
    • CASP3
    • Caspase 3 apoptosis related cysteine protease
    • Caspase 3 p12 subunit
    • Caspase 3 p17 subunit
    • Caspase3
    • CPP 32
    • CPP32
    • CPP32B
    • Cysteine protease CPP32
    • Human cysteine protease CPP32 isoform alpha mRNA complete cds
    • ICE3
    • LICE
    • PARP cleavage protease
    • SCA 1
    • SCA1
    • SREBP cleavage activity 1
    • Yama protein
    see all

Applications

Our Abpromise guarantee covers the use of ab39383 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent concentration.

Protocols

References for Caspase 3 Assay Kit (Fluorometric) (ab39383)

This product has been referenced in:
  • Burton TR  et al. BNIP3 acts as transcriptional repressor of death receptor-5 expression and prevents TRAIL-induced cell death in gliomas. Cell Death Dis 4:e587 (2013). Read more (PubMed: 23579274) »
  • Mejías R  et al. Long term biotransformation and toxicity of dimercaptosuccinic acid-coated magnetic nanoparticles support their use in biomedical applications. J Control Release 171:225-33 (2013). Read more (PubMed: 23906866) »

See all 3 Publications for this product

Product Wall



I am afraid that your protease inhibitors might interefere with the assay. Since Caspase is a cysteine protease it is probably inhibited by most protease inhibitor cocktails. The assay is based on the catalytic clevage of the substrate DEVD-A...

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Thank you for your inquiry. This kit will detect active Caspase-3 from samples. The procasepase-3 (inactive) is proteolytically processed into the active Caspase-3. I hope this information helps.  Please contact us with any other questions.

Unfortunately, this is proprietary information.

Thank you for your enquiry and your interest in our products. The sensitivity range for the fluorometric substrate (K105) is in the tenths of a µM, while the sensitivity of the colorimetric substrates (K106) is in the single digit µM. These kits ca...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"