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

Submit an Abreview Q&A (2)Specific References (2)

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: Other species, Mammals
  • 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.

  • Tested applications
    Suitable for: Flow Cytmore details
  • Platform
    Flow

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    500 X TF5-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

Applications

Our Abpromise guarantee covers the use of ab112130 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
Flow Cyt Use at an assay dependent concentration.

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.

Protocols

References

This product has been referenced in:
  • Chang HW  et al. Withaferin A Induces Oxidative Stress-Mediated Apoptosis and DNA Damage in Oral Cancer Cells. Front Physiol 8:634 (2017). Read more (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). Read more (PubMed: 27827950) »

See all 2 Publications for this product

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

Customer reviews and Q&As

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2 reviews or Q&A

10
Votes: Highest First

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?

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 anyt...

Read More

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!

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.

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

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