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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 Studies more 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
        ApopainApopain precursor CASP 3
        CASP3Caspase 3 apoptosis related cysteine proteaseCaspase 3 p12 subunitCaspase 3 p17 subunitCaspase3CPP 32CPP32CPP32BCysteine protease CPP32Human cysteine protease CPP32 isoform alpha mRNA complete cdsICE3LICEPARP cleavage proteaseSCA 1SCA1SREBP cleavage activity 1Yama 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 Notes
    Functional Studies FuncS

    Protocols

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

    This product has been referenced in:
    • Lee YJ  et al. Apoptotic cell instillation after bleomycin attenuates lung injury through hepatocyte growth factor induction. Eur Respir J 40:424-35 (2012). Read more (PubMed: 22441736) »

    See 1 Publication for this product

    Publishing research using ab39383 ? Please let us know so that we can cite the reference in this datasheet

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    Submit a Question for ab39383

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    Will this kit detect full length caspase 3 or cleaved caspase 3?

    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.    

    Thank you for your reply. Our user wants know detail component of cell lysis buffer.

    Unfortunately, this is proprietary information.

    i need senstivity range and Species Reactivity for ab 39401 and AB 3983. i need these kit for rats.

    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 can d...

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