Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Microplate reader, Fluor. microscope, Flow cyt.
- Sample type: Adherent cells, Suspension cells
Product nameCaspase-1 (active) Staining Kit - Green Fluorescence
See all Caspase-1 kits
Sample typeAdherent cells, Suspension cells
Species reactivityReacts with: Mammals, Other species
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.
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).
PlatformMicroplate reader, Fluor. microscope, Flow cyt.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 25 tests 500X Hoechst Stain 1 x 100µl 500X Propidium Iodide 1 x 100µl FAM-YVAD-FMK 1 vial Washing Buffer 1 x 100ml
RelevanceCaspases 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.
- CASP1 nirs variant 1
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.
ab219935 has been referenced in 2 publications.
- Long Y et al. ROS-induced NLRP3 inflammasome priming and activation mediate PCB 118- induced pyroptosis in endothelial cells. Ecotoxicol Environ Saf 189:109937 (2020). PubMed: 31785945
- 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 . PubMed: 28246166