Overview

  • Product name
    Caspase 1 (active) Staining Kit - Green Fluorescence
    See all Caspase 1 kits
  • Detection method
    Fluorescent
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
    Cell-based
  • Species reactivity
    Reacts with: Other species, Mammals
  • Product overview

    Caspase 1 (active) Staining Kit - Green Fluorescence (ab219935) is a sensitive fluorometric assay to measure caspase 1 activation in live cells. The assay uses FAM-YVAD-FMK, which binds irreversibly to active caspase 1 in stimulated cells. The fluorescent intensity of the FAM-YVAD-FMK signal is proportional to the amount of active caspase 1 and can be easily detected at Ex/Em = 490/525 nm by fluorescence microscopy, flow cytometer, or fluorescent microplate reader.

  • Notes

    Caspase activity assay kits are based on fluorescent inhibitors of caspases. These inhibitors are cell permeable and non-cytotoxic. Once inside the cell, the caspase inhibitors bind covalently to the active caspases. Caspase-1 is primarily involved in the activation of pro-inflammatory cytokines and the process of pyropotosis. It has been proven that caspase 1 has substrate selectivity for the peptide sequence Tyr-Val-Ala-Asp (YVAD).

  • Tested applications
    Suitable for: FM, Flow Cytmore details

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 25 tests
    500X Hoechst Stain 1 vial
    500X Propidium Iodide 1 vial
    FAM-YVAD-FMK 1 vial
    Washing Buffer 1 x 100ml
  • Research areas
  • Relevance
    Caspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis. The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a2b2 tetramer to form the active enzyme. Caspase 1 is similar to the cell death gene CED3 of C. elegans and regulates multiple proinflammatory cytokines, including Interleukin 1b and interferon-gamma-inducing factor. Caspase 1 plays a role in down stream of Caspase 8 which is involved in Fas-mediated apoptosis.
  • Cellular localization
    Cytoplasmic
  • Alternative names
    • CASP1
    • CASP1 nirs variant 1
    • Caspase 1 apoptosis related cysteine peptidase
    • caspase 1, apoptosis-related cysteine peptidase (interleukin 1, beta, convertase)
    • ICE
    • ICE-like protease
    • IL1 B convertase
    • IL1 beta converting enzyme
    • IL1BC
    • IL1BCE
    • Interleukin 1 B converting enzyme
    • Interleukin 1 beta convertase
    • P45
    see all

Applications

Our Abpromise guarantee covers the use of ab219935 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
FM Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

Images

  • Detection of active Caspase 1 in Jurkat cells. Jurkat cells (3 x 105 cells/100 μL/well) were either untreated (control) or treated with 1 μM staurosporine for 3 hours. Cells were incubated with FAM-YVAD-FMK for 1 hour at 37°C. The fluorescent signal was measured at Ex/Em = 490/525 nm (cut off at 515 nm) with a FlexStation microplate reader (Molecular Devices) using bottom read mode.

  • Active caspase 1 staining in Jurkat cells. cells (3 x 105 cells/100 μL/well) were either untreated (A) or treated with 1 μM staurosporine for 3 hours (B). Cells were incubated with FAM-YVAD-FMK for 1 hour at 37°C. Increase in fluorescent intensity was observed using a fluorescence microscope with a FITC channel.

Protocols

References

This product has been referenced in:
  • Parzych K  et al. Differential role of pannexin-1/ATP/P2X7 axis in IL-1ß release by human monocytes. FASEB J 31:2439-2445 (2017). Functional Studies ; Human . Read more (PubMed: 28246166) »

See 1 Publication for this product

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

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