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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).
|500X Hoechst Stain||1 vial|
|500X Propidium Iodide||1 vial|
|Washing Buffer||1 x 100ml|
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.
|FM||Use at an assay dependent concentration.|
|Functional Studies||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.|
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.