Cell Meter™ Generic Fluorimetric Caspase Activity Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
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International | See distributors |
Bulk request | Inquire |
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Shipping | Standard overnight for United States, inquire for international |
Spectral properties
Correction Factor (280 nm) | 0.09 |
Extinction coefficient (cm -1 M -1) | 75000 |
Excitation (nm) | 503 |
Emission (nm) | 525 |
Quantum yield | 0.9 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Related products
Overview | ![]() ![]() |
See also: Caspases
Correction Factor (280 nm) 0.09 | Extinction coefficient (cm -1 M -1) 75000 | Excitation (nm) 503 | Emission (nm) 525 | Quantum yield 0.9 |
Our Cell Meter™ assay kits are a set of tools for monitoring cell viability. There are a variety of parameters that can be used for monitoring cell viability. This particular kit is designed to monitor cell apoptosis through measuring generic caspases (caspase-1, -3, -4, -5, -6, -7, -8 and -9) activation in living cells. Caspases activation is widely accepted as a reliable indicator for cell apoptosis. Most caspases have substrate selectivity for the peptide sequence Val-Ala-Asp (VAD). This kit uses TF2-VAD-FMK as a fluorogenic indicator for most caspase activity. TF2-VAD-FMK is cell permeable, nontoxic, and irreversibly binds to activated casepase-1, -3, -4, -5, -6, -7, -8 and -9 in apoptotic cells. Once bound to caspases, the green fluorescent reagent is retained within the cell. The binding event prevents the caspases from further catalysis but will not stop apoptosis from proceeding. The reagent will start to react with active caspase enzymes within 15 minutes of addition to the media. The kit provides all the essential components with an optimized assay protocol. It is used for the quantification of most activated caspases activities in apoptotic cells, or for screening caspases inhibitors. The green label allows for direct detection of activated caspases in apoptotic cells by flow cytometry.
Platform
Flow cytometer
Excitation | 488 nm laser |
Emission | 530/30 nm filter |
Instrument specification(s) | FITC channel |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with test compounds at a density of 5 × 105 to 1 × 106 cells/mL
- Add 1 µL of 500X TF2-VAD-FMK into 0.5 mL of cell solution
- Incubate the cells in a 37°C, 5% CO2 incubator for 1 - 4 hours
- Pellet the cells and resuspend the cells in 0.5 mL of assay buffer or growth medium
- Analyze cells using flow cytometer with 530/30 nm filter (FITC channel)
Important notes
Thaw all the kit components at room temperature before starting the experiment.
SAMPLE EXPERIMENTAL PROTOCOL
- For each sample, prepare cells in 0.5 mL warm medium or buffer of your choice at a density of 5 × 105 to 1 × 106 cells/mL. Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density for apoptosis induction.
- Treat cells with test compounds for a desired period of time to induce apoptosis, and create positive and negative controls.
- Add 1 µL of 500X TF2-VAD-FMK (Component A) into the treated cells.
- Incubate the cells in a 37°C, 5% CO2 incubator for 1 - 4 hours. Note: For adherent cells, gently lift the cells with 0.5 mM EDTA to keep the cells intact, and wash the cells once with serum-containing media prior to incubation with TF2-VAD-FMK. The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.
- Wash and spin the cells twice. Resuspend the cells in 0.5 mL of Assay Buffer (Component B) or growth medium. Note: TF2-VAD-FMK is fluorescent; therefore it is important to wash out any unbound reagent to remove the background.
- If desired, label the cells with a DNA stain (such as propidium iodide or 7-AAD for dead cells).
- If desired, fix cells.
- Monitor the fluorescence intensity using a flow cytometer wih 530/30 nm filter (FITC channel). Gate on the cells of interest, excluding debris.
Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Correction Factor (280 nm) | 0.09 |
Extinction coefficient (cm -1 M -1) | 75000 |
Excitation (nm) | 503 |
Emission (nm) | 525 |
Quantum yield | 0.9 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Correction Factor (280 nm) |
Cell Meter™ Generic Fluorimetric Caspase Binding Assay Kit *Red Fluorescence Optimized for Flow Cytometry* | 649 | 664 | 250000 | 0.027 |
Images

Figure 1. Detection of caspase activity using Cell Meter™ Generic Fluorometric Caspase Activity Assay Kit in Jurkat cells. TF2-VAD-FMK fluorescence intensity was induced with the addition of camptothecin in Jurkat cells. Jurkat cells were treated without (Blue) or with 20 µM camptothecin (Red) in a 37 °C, 5% CO2 incubator for 4-5 hours, and then dye loaded with TF2-VAD-FMK for 1 hour. Response was recorded using BD FACSCalibur flow cytomter using FL-1 channel.
Citations
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Extract of Juniperus indica Bertol synergizes with cisplatin to inhibit oral cancer cell growth via repression of cell cycle progression and activation of the caspase cascade
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Journal: Molecules (2020): 2746
Promotion of osteointegration under diabetic conditions by tantalum coating-based surface modification on 3-dimensional printed porous titanium implants
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Application notes
FAQ
Are inflammasomes and caspase-1 related?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?