Caspase 3 Assay Kit (Colorimetric) (ab39401)
- Product nameCaspase 3 Assay Kit (Colorimetric)See all Caspase 3 kits ...
- Tests100 x 1 assay
- Sample typeTissue Extracts, Cell Lysate
- Assay typeEnzyme activity
- Assay time2h 00m
- Product overview
Abcam's Caspase 3 Assay Kit (Colorimetric) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. The assay is based on spectrophotometric detection of the chromophore p-nitroaniline (p-NA) after cleavage from the labeled substrate DEVD-pNA. The p-NA light emission can be quantified using a spectrophotometer or a microtiter plate reader at 400- or 405 nm. Comparison of the absorbance of p-NA from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase 3 activity.
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Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells.
If you need more DEVD-pNA substrate, it is available for purchase as ab171038.
- Tested applicationsFunctional Studies more details
- Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests 2X Reaction Buffer 4 x 2ml Cell Lysis Buffer 1 x 100ml DEVD-pNA 1 x 500µl Dilution Buffer 1 x 100ml DTT Green 1 x 400µl
- 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
- Apopain precursor
- CASP 3
- Caspase 3 apoptosis related cysteine protease
- Caspase 3 p12 subunit
- Caspase 3 p17 subunit
- CPP 32
- Cysteine protease CPP32
- Human cysteine protease CPP32 isoform alpha mRNA complete cds
- PARP cleavage protease
- SCA 1
- SREBP cleavage activity 1
- Yama protein
Our Abpromise guarantee covers the use of ab39401 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Caspase 3 Assay Kit (Colorimetric) images
H9c2 cells were either untreated (control) or treated with 2µg/ml of recombinant lipocalin-2 (Lcn2) for 24h. Cell lysates were assayed for caspase-3 activity (n = 3, *, p<0.05).
Image obtained from Xu G et al., JBC, 2012 Feb 11;28(7):4808-17
References for Caspase 3 Assay Kit (Colorimetric) (ab39401)
This product has been referenced in:
- Helmer RA et al. Role of helicase-like transcription factor (hltf) in the G2/m transition and apoptosis in brain. PLoS One 8:e66799 (2013). Read more (PubMed: 23826137) »
- Xu G et al. Lipocalin-2 induces cardiomyocyte apoptosis by increasing intracellular iron accumulation. J Biol Chem 287:4808-17 (2012). Read more (PubMed: 22117066) »