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    products/assay-kits/cleaved-caspase-3-staining-kit-fitc-ab65613.pdf

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Cell Biology Apoptosis Intracellular Caspases etc Caspases
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Cleaved Caspase-3 Staining Kit (FITC) (ab65613)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1) Submit a question References (21)

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Functional Studies - Caspase-3 (active) FITC Staining Kit (ab65613)
  • Functional assays: Caspase-3 (active) FITC Staining Kit (ab65613)

Key features and details

  • Assay type: Enzyme activity
  • Platform: Microplate reader, Fluor. microscope, Flow cyt.
  • Assay time: 2 hr
  • Sample type: Adherent cells, Suspension cells

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Overview

  • Product name

    Cleaved Caspase-3 Staining Kit (FITC)
    See all Caspase-3 kits
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Enzyme activity
  • Assay time

    2h 00m
  • Product overview

    Cleaved Caspase-3 Staining Kit (FITC) ab65613 provides a convenient means for sensitive detection of activated Caspase-3 in living cells.


    The cleaved caspase 3 assay utilizes the Caspase-3 inhibitor, DEVD-FMK, conjugated to FITC (FITC-DEVD-FMK) as a marker. FITC-DEVD-FMK is cell permeable, non-toxic, and irreversibly binds to activated Caspase-3 in apoptotic cells. The FITC label allows detection of activated / cleaved caspase-3 in apoptotic cells directly by fluorescence microscopy, flow cytometry, or fluorescence plate reader.

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K183 CaspGLOW™ Fluorescein Active Caspase-3 Staining Kit. K183-100 is the same size as the 100 test size of ab65613.

    Other caspase and apoptosis assays

    Review the full set of caspase assays, or the apoptosis assay and apoptosis marker guide.

  • Platform

    Microplate reader, Fluor. microscope, Flow cyt.

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    FITC-DEVD-FMK 1 x 100µl
    Wash Buffer 2 x 100ml
    Z-VAD-FMK 1 x 10µl
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Caspases
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Other proteases
    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • Caspase Assays Kits
    • Caspase 3 assay kits
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Caspases
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
  • Relevance

    The 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 (e.g. 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 localization

    Cytoplasmic
  • Alternative names

    • A830040C14Rik
    • Apopain
    • Apopain precursor
    • CASP 3
    • CASP-3
    • CASP3
    • CASP3_HUMAN
    • Casp3a
    • Caspase 3 apoptosis related cysteine protease
    • Caspase 3 p12 subunit
    • Caspase 3 p17 subunit
    • Caspase 3, apoptosis-related cysteine peptidase
    • Caspase 3, apoptosis-related cysteine protease
    • Caspase 3, apoptosis-related cysteine protease a
    • Caspase-3 subunit p12
    • Caspase3
    • CC3
    • CP32B
    • CPP 32
    • CPP-32
    • CPP32
    • CPP32B
    • Cysteine protease CPP32
    • EC 3.4.22.56
    • ICE3
    • LICE
    • mldy
    • OTTHUMP00000165052
    • OTTHUMP00000165053
    • OTTHUMP00000165054
    • PARP cleavage protease
    • Procaspase3
    • Protein Yama
    • SCA 1
    • SCA-1
    • SCA1
    • SREBP cleavage activity 1
    • Yama
    • Yama protein
    see all

Associated products

  • Related Products

    • Z-D(OMe)E(OMe)VD(OMe)-FMK, Cell permeable caspase-3 inhibitor (ab120488)
    • Z-VDVAD-FMK, caspase-2 inhibitor (ab142035)
    • Q-DEVD-OPh, caspase-3 inhibitor (ab142037)
    • PAC-1, caspase-3 activator (ab142074)

Images

  • Functional Studies - Caspase-3 (active) FITC Staining Kit (ab65613)
    Functional Studies - Caspase-3 (active) FITC Staining Kit (ab65613)Mart?nez-Torres, Ana Carolina et al., BMC cancer?vol. 18,1 13., Fig 6, doi:10.1186/s12885-017-3954-5

    Caspase-3 activation was measured by flow cytometry, using ab65613, in cells that were left alone or pretreated with QVD.oph and then treated with I-CRP (1.25 U/ml) or a drug that induces DNA strand breaks (100 µM) for 24 h, data was then analyzed and graphed.

  • Functional assays: Caspase-3 (active) FITC Staining Kit (ab65613)
    Functional assays: Caspase-3 (active) FITC Staining Kit (ab65613)

    Caspase-3 in Jurkat cells following 24 hour exposure to 2 uM Camptothecin (CPT; ab120115) with or without 20 nM caspase inhibitor Z-VAD(OMe)-FMK (zVAD; ab120487). Background signal subtracted, duplicates; +/- SD.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (21)

Publishing research using ab65613? Please let us know so that we can cite the reference in this datasheet.

ab65613 has been referenced in 21 publications.

  • Di Franco S  et al. CHK1 inhibitor sensitizes resistant colorectal cancer stem cells to nortopsentin. iScience 24:102664 (2021). PubMed: 34169240
  • Zheng JJ  et al. Novel role of PAF1 in attenuating radiosensitivity in cervical cancer by inhibiting IER5 transcription. Radiat Oncol 15:131 (2020). PubMed: 32471508
  • Ding D  et al. Some Ototoxic Drugs Destroy Cochlear Support Cells Before Damaging Sensory Hair Cells. Neurotox Res 37:743-752 (2020). PubMed: 31997155
  • Poholek CH  et al. Noncanonical STAT3 activity sustains pathogenic Th17 proliferation and cytokine response to antigen. J Exp Med 217:N/A (2020). PubMed: 32697822
  • Eberle-Singh JA  et al. Effective Delivery of a Microtubule Polymerization Inhibitor Synergizes with Standard Regimens in Models of Pancreatic Ductal Adenocarcinoma. Clin Cancer Res 25:5548-5560 (2019). PubMed: 31175095
View all Publications for this product

Customer reviews and Q&As

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Use of Caspase 3 (Active) FITC Staining Kit for invertebrate tissues

Below average Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Attempted to use this stain for an in-tact invertebrate organ - specifically, the light organ of the Hawaiian Bobtail squid, Euprymna scolopes. The stain was easy to use, and worked beautifully on some surface-level tissues of the animal. E.g, apoptotic cells with caspase 3 in the tentacle/arms were clearly visible. However, the light organ - which is in the squid body cavity and overlaid on one side with the gills and the other with the funnel - was not compatible with this stain, hence the 2-star review. The following adaptations of the prescribed protocol were attempted:

1) Whole animals incubated with the dye for 30/60 minutes at 37, washed with the wash buffer.
2) Ventrally-dissected animals (light organ exposed) incubated with the dye for 30 and 60 minutes at 37, then washed with wash buffer.
3) Completely removed light organs incubated with the dye for 30 and 60 minutes at 37, then washed with wash buffer.
4) All of the above repeated, but permeabilized by 1 and 2% Triton X 100 for 30 minutes before staining.

Apoptosis in the light organ tissues was always verified before caspase staining by staining with acridine orange. For some reason, however, the FITC-conjugated inhibitor peptide was never observed to penetrate into the light organ. In the picture attached, the arms of the light organ extending outwards showed signs of apoptotic cell death when examined by arcridine orange (image not shown). However, nothing was visible with the caspase dye (image shown).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Aug 08 2018

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