CellTox™ Green Cytotoxicity Assay
Real-Time Cell Death Assay With Multiplexing Compatibility
- Accurate cytotoxicity determination in exposures out to 72 hours
- Flexible protocol allows kinetic analysis or endpoint determination
- Multiplex with luminescent assays to obtain more data per well
Catalog Number:
Size
Catalog Number: G8741
Catalog Number: G8742
Catalog Number: G8743
Catalog Number: G8731
Monitoring Cell Death in Real Time Using the CellTox™ Green Cytotoxicity Assay
Cytotoxicity assays based on detection of biomarkers released into the media can underestimate cytotoxicity in long-term exposures of 72 hours or more because of limited stability of the biomarker detected. The CellTox™ Green Cytotoxicity Assay provides an easy, fast and accurate method to determine toxic effects during or after long-term exposure of cells in culture. CellTox Green can be combined with other methods in multiplex assays to determine mechanism of toxicity, and is easily scalable from 96- to 1536-well plate formats.
Learn how the CellTox™ Green Cytotoxicity Assay can be used to assess drug toxicity in organoids.
CellTox™ Green Dye binds DNA of cells with impaired membrane integrity.
Perform Kinetic Cytotoxicity Measures to Determine Onset of Toxicity
Dose- and exposure-dependent increases in cytotoxicity
In the experiment shown here, bortezomib caused appreciable cytotoxicity in K562 cells between the 4- and 24-hour time points. Additive fluorescence at later time points indicates continued loss of membrane integrity occurring as a function of cytotoxicity against remaining cells. The CellTox™ Green Dye was added at seeding.
Multiplex with Cell Viability Assays for More Informative Data per Well
Measure Cytotoxicity and Viability
Here, CellTox™ Green Dye was mixed with K562 Cells, which were plated, then dosed with compound. CellTiter-Glo® Cell Viability Assay Reagent was added at the end of the 72-hour exposure and luminescence (viability) measured. These two inverse measures of cell health resulted in EC50 agreement.
Determine Appropriate Timing of Caspase Assays
CellTox™ Green can be used in kinetic mode to determine the onset of cytotoxicity followed by multiplexing with the Caspase-Glo® 3/7 Assay in the same sample well to confirm caspase-3/7 activation at the appropriate time.
Differentiate Between Cytotoxic and Cytostatic Effects
In this example, K562 cells were exposed to Methotrexate, an anti-proliferative compound, for 72 hours. CellTox™ Green Dye was added to determine cytotoxicity, then CellTiter-Glo® Reagent was added to the same sample well for viability determination. A viability assay alone would have misreported the compound as cytotoxic due to less cells in the treated sample compared to the non-treated control, which continued to proliferate. Multiplexing with a cytotoxicity assay reveals no change in signal as a result of compromised membrane integrity, indicating growth inhibition and not cytotoxicity.
After several months trying to develop a robust cytotoxicity cell-based assay, we always observed high variability on the upper asymptote of the dose-response curve. Based on Promega’s recommendations, we replaced our previous cytotoxicity detection reagent by the CellTox™ Green Cytotoxicity reagent, which directly solved our issues. The cell-based assay is now fully validated with this reagent in our company.
Dr. Fabian Vandermeers, Sr R&D Technical Leader, Quality Assistance S.A.
Protocols
Complete Protocol
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
|
Lysis Solution |
G182B | 1 × 0.5ml |
|
Assay Buffer |
G610A | 1 × 10ml |
CellTox™ Green Dye, 1,000X |
G873A | 1 × 20μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size |
|---|---|---|
|
Lysis Solution |
G182B | 1 × 0.5ml |
|
Assay Buffer |
G610B | 1 × 50ml |
CellTox™ Green Dye, 1,000X |
G873A | 5 × 20μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size |
|---|---|---|
|
Lysis Solution |
G182B | 1 × 0.5ml |
|
Assay Buffer |
G610B | 2 × 50ml |
CellTox™ Green Dye, 1,000X |
G873B | 1 × 200μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size |
|---|---|---|
CellTox™ Green Dye, 1,000X |
G873B | 1 × 200μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
Resources
Articles
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PLos ONE
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