Generic Caspase Activity Assay Kit (Fluorometric - Green) (ab112130)

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Flow Cytometry - Generic Caspase Activity Assay Kit (Fluorometric - Green) (ab112130)

    Key features and details

    • Assay type: Direct
    • Detection method: Fluorescent
    • Platform: Flow cytometer
    • Sample type: Adherent cells, Suspension cells

    Overview

    • Product name

      Generic Caspase Activity Assay Kit (Fluorometric - Green)
      See all Caspase kits

    • Detection method

      Fluorescent

    • Sample type

      Adherent cells, Suspension cells

    • Assay type

      Direct

    • Species reactivity

      Reacts with: Mammals, Other species

    • Product overview

      Abcam's Activity Assay kits are a set of tools for monitoring cellular functions. The activation of caspase is widely accepted as a reliable indicator for cell apoptosis. Most caspases have substrate selectivity for the peptide sequence Val-Ala-Asp (VAD). ab112130 uses TF2-VAD-FMK as a fluorescent indicator for most caspase activities. The cell permeable and nontoxic TF2-VAD-FMK irreversibly binds to activated caspase-1, -3, -4, -5, -6, -7, -8 and -9 in apoptotic cells. Once bound to caspases, the fluorescent reagent is retained inside the cell. The binding event prevents the caspases from further catalysis but will not stop apoptosis from proceeding. Within 15 minutes incubation, it starts to react with active caspase enzymes.


      ab112130 provides all the essential components with an optimized assay protocol. It is designed to detect cell apoptosis by measuring generic activation of caspases (caspase-1, -3, -4, -5, -6, -7, -8 and -9) in live cells.


      ab112130 is used for either the quantification of most activated caspase activities in apoptotic cells or screening of caspase inhibitors. TF2-VAD-FMK, the green label reagent, allows for direct detection of activated caspases in apoptotic cells by a flow cytometer at Ex/Em = 488/520 nm.

      Visit our FAQs page for tips and troubleshooting.

    • Notes

      ab112130 should be stored dessicated.

      Other caspase and apoptosis assays

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

    • Platform

      Flow cytometer

    Properties

    • Storage instructions

      Store at -20°C. Please refer to protocols.
    • Components 100 tests
      500 X TF2-VAD-FMK 1 x 100µl
      500X Propidium Iodide 1 x 100µl
      Assay Buffer 1 x 50ml

    • Research areas

    • Relevance

      Caspases are members of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce 2 subunits, large and small, that dimerize to form the active enzyme.

    • Cellular localization

      Cytoplasmic

    Images

    • Flow Cytometry - Generic Caspase Activity Assay Kit (Fluorometric - Green) (ab112130)
      Flow Cytometry - Generic Caspase Activity Assay Kit (Fluorometric - Green) (ab112130)
      The increase in TF2-VAD-FMK (Component A) fluorescence intensity with the addition of Camptothecin in Jurkat cells.
      Jurkat cells were untreated (Blue) or with 20 µM camptothecin (Pink) in a 37 °C, 5 % CO2 incubator for 4-5 hours, and then dye loaded with TF2-VAD-FMK for 1 hour.

    References (6)

    Publishing research using ab112130? Please let us know so that we can cite the reference in this datasheet.

    ab112130 has been referenced in 6 publications.

    • Petroni G  et al. Clarithromycin inhibits autophagy in colorectal cancer by regulating the hERG1 potassium channel interaction with PI3K. Cell Death Dis 11:161 (2020). PubMed: 32123164
    • Pillozzi S  et al. Peptides and small molecules blocking the CXCR4/CXCL12 axis overcome bone marrow-induced chemoresistance in acute leukemias. Oncol Rep 41:312-324 (2019). PubMed: 30365110
    • Huang HW  et al. Sinularin Selectively Kills Breast Cancer Cells Showing G2/M Arrest, Apoptosis, and Oxidative DNA Damage. Molecules 23:N/A (2018). PubMed: 29642488
    • Nicholson J  et al. E3 Ligase cIAP2 Mediates Downregulation of MRE11 and Radiosensitization in Response to HDAC Inhibition in Bladder Cancer. Cancer Res 77:3027-3039 (2017). PubMed: 28363998
    • Chang HW  et al. Withaferin A Induces Oxidative Stress-Mediated Apoptosis and DNA Damage in Oral Cancer Cells. Front Physiol 8:634 (2017). PubMed: 28936177
    • Chen CY  et al. Tenuifolide B from Cinnamomum tenuifolium Stem Selectively Inhibits Proliferation of Oral Cancer Cells via Apoptosis, ROS Generation, Mitochondrial Depolarization, and DNA Damage. Toxins (Basel) 8:N/A (2016). PubMed: 27827950

    Customer reviews and Q&As

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    Question

    However, If i fix the cell sample using 70% alcohol such as cell
    stained by PI to runing cell cycle then the sample just can store at 4
    oC for 24hour, right?

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    Answer

    Thank you for your reply.

    You are correct that you can your cells for up to 24hours at 4C after they have been fixed. After the cells have been fixed. The kit is intended to be used on live cells.

    Please let me know if there is anything else I can help you with

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    Question

    Inthe protocolofthis productionwhichpage 8, the F step. For cell fixation, how long can it bestored at -20 or -80 oC?
    I need the important information, please help me.
    thanks a lot!

    Read More
    Answer

    Thank you forcontacting Abcam.

    Youcan store your samples at 4C for up to 24hours, but you cannot freeze them.

    Please let me know if there is anything else I can help you with.

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