Live or Dead™ Cell Viability Assay Kit *Red/Blue Dual Fluorescence*
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Spectral properties
Excitation (nm) | 612 |
Emission (nm) | 630 |
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 |
Alternative formats
Live or Dead™ Cell Viability Assay Kit *Green/Red Dual Fluorescence* |
Related products
Overview | ![]() ![]() |
See also: Live or Dead Cell Viability Assays
Excitation (nm) 612 | Emission (nm) 630 |
This Live or Dead™ Cell Viability Assay Kit uses two fluorescent indicators: Cellbrite™ Red (Ex/Em = 613/631 nm) for labeling viable cells and a cell-impermeable DNA-binding dye Nuclear Blue™ DCS1 (Ex/Em = 360/450 nm) for labeling dead cells with damaged membranes. Cells grown in black-wall plates can be stained and quantified in less than two hours. The assay is more robust and accurate than the other viability assays. It can be readily adapted for a wide variety of fluorescence platforms such as microplate assays, fluorescence microscopes, and flow cytometry. The kit provides all the essential components with an optimized assay protocol. It is suitable for both proliferating and non-proliferating cells (either suspension or adherent cells).
Platform
Fluorescence microscope
Excitation | Cy5 filter (alive), DAPI filter (dead) |
Emission | Cy5 filter (alive), DAPI filter (dead) |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 610, 360 nm |
Emission | 650, 450 nm |
Cutoff | 630, 420 nm |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Bottom read mode |
Components
Example protocol
AT A GLANCE
Protocol Summary
- Prepare cells with test compounds
- Add dye-working solution
- Incubate at room temperature or 37°C for 30 minutes to 1 hour
- Monitor fluorescence intensity (bottom read mode) at Ex/Em = 610/650 nm (Cutoff = 630 nm, Red) and Ex/Em = 360/450 nm (Cutoff =420 nm, Blue) or fluorescence microscope with Cy5 channel (live) and DAPI channel (dead)
Important Note
Thaw all the kit components at room temperature before starting the experiment.
CELL PREPARATION
For guidelines on cell sample preparation, please visit:
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
PREPARATION OF WORKING SOLUTION
Add 5 µL of 200X Cellbrite™ Red (Component A) and 5 µL of 200X Nuclear Blue™ DCS1 (Component C) into 1 mL of Assay Buffer (Component B) and mix well to make dye-working solution. This dye-working solution is stable for at least 1 hour at room temperature.
Note: As the optimal staining conditions may vary depending on different cell types, it’s recommended to determine the appropriate concentration of Component A and C individually.
SAMPLE EXPERIMENTAL PROTOCOL
- Prepare cells according to the standard protocol. Note: We treated HeLa cells with staurosporine (SS) for 4 hours at 37ºC to induce cell apoptosis. See Figure 1 for details.
- Replace growth medium with 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of dye-working solution.
- Incubate the dye-working solution plate at room temperature or 37°C for 30 minutes to 1 hour, protected from light.
- Wash cells with HHBS, PBS or buffer of your choice twice.
- Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Assay Buffer (Component B) into the cells.
- Monitor the fluorescence signal under a fluorescence microscope with Texas Red or Cy5 filter for live cells, and DAPI filter for dead cells. The fluorescence intensity can also be analyzed with a fluorescence microplate reader (bottom read mode) at Ex/Em = 610/650 nm (Cutoff = 630 nm, Red) and Ex/Em = 360/450 nm (Cutoff =420 nm, Blue).
Images

Figure 1. Fluorescence images of HeLa cells labeled with Live or Dead™ Cell Viability Assay Kit *Dual Fluorescence* (Cat#22788). HeLa cells at 100,000 cells/well/100 µL were seeded overnight in a 96-well black wall/clear bottom plate. Cells were treated with 0-1 µM staurosporine (SS) at 37ºC for 4 hours (A-D), or fixed in ethanol (E), then incubated with dye-loading solution for 1 hour. The fluorescence signal was measured using a fluorescence microscope with Texas Red or Cy5 filter for viable cells (Red) and DAPI filter for necrotic cells (Blue), respectively. (F) The corresponding fluorescence signal were measured using a FlexStation® microplate reader (Molecular Devices) with bottom read mode at Ex/Em= 610/650 (cutoff=630 nm, Red) and Ex/Em=360/450 (cutoff=420 nm, Blue), respectively.
Citations
View all 20 citations: Citation Explorer
Developing a Novel Platelet-Rich Plasma-Laden Bioadhesive Hydrogel Contact Lens for the Treatment of Ocular Surface Chemical Injuries
Authors: Soykan, Merve Nur and Altug, Burcugul and Bas, Harun and Ghorbanpoor, Hamed and Avci, Huseyin and Eroglu, Sertac and Sengel, Sultan Butun and Sariboyaci, Ayla Eker and Bagis, Sibel Gunes and Uysal, Onur and others,
Journal: Macromolecular Bioscience (2023): 2300204
Authors: Soykan, Merve Nur and Altug, Burcugul and Bas, Harun and Ghorbanpoor, Hamed and Avci, Huseyin and Eroglu, Sertac and Sengel, Sultan Butun and Sariboyaci, Ayla Eker and Bagis, Sibel Gunes and Uysal, Onur and others,
Journal: Macromolecular Bioscience (2023): 2300204
A Multifunctional Sateen Woven Dressings for Treatment of Skin Injuries
Authors: Ozel, Ceren and Apaydin, Elif and Sariboyaci, Ayla Eker and Tamayol, Ali and Avci, Huseyin
Journal: Colloids and Surfaces B: Biointerfaces (2023): 113197
Authors: Ozel, Ceren and Apaydin, Elif and Sariboyaci, Ayla Eker and Tamayol, Ali and Avci, Huseyin
Journal: Colloids and Surfaces B: Biointerfaces (2023): 113197
Ultrasound and microbubbles (USMB) potentiated doxorubicin penetration and distribution in 3D breast tumour spheroids
Authors: Misra, Rahul and Rajic, Mathew and Sathiyamoorthy, Krishnan and Karshafian, Raffi
Journal: Journal of Drug Delivery Science and Technology (2020): 102261
Authors: Misra, Rahul and Rajic, Mathew and Sathiyamoorthy, Krishnan and Karshafian, Raffi
Journal: Journal of Drug Delivery Science and Technology (2020): 102261
Functional imaging of neuronal activity of auditory cortex by using Cal-520 in anesthetized and awake mice
Authors: Li, Jingcheng and Zhang, Jianxiong and Wang, Meng and Pan, Junxia and Chen, Xiaowei and Liao, Xiang
Journal: Biomedical Optics Express (2017): 2599--2610
Authors: Li, Jingcheng and Zhang, Jianxiong and Wang, Meng and Pan, Junxia and Chen, Xiaowei and Liao, Xiang
Journal: Biomedical Optics Express (2017): 2599--2610
NINJ2--A novel regulator of endothelial inflammation and activation
Authors: Wang, Jingjing and Fa, Jingjing and Wang, Pengyun and Jia, Xinzhen and Peng, Huixin and Chen, Jing and Wang, Yifan and Wang, Chenhui and Chen, Qiuyun and Tu, Xin and others, undefined
Journal: Cellular Signalling (2017)
Authors: Wang, Jingjing and Fa, Jingjing and Wang, Pengyun and Jia, Xinzhen and Peng, Huixin and Chen, Jing and Wang, Yifan and Wang, Chenhui and Chen, Qiuyun and Tu, Xin and others, undefined
Journal: Cellular Signalling (2017)
Influence of hypothermia and subsequent rewarming upon leukocyte-endothelial interactions and expression of Junctional-Adhesion-Molecules A and B
Authors: Bogert, Nicolai V and Werner, Isabella and Kornberger, Angela and Meybohm, Patrick and Moritz, Anton and Keller, Till and Stock, Ulrich A and Beiras-Fern, undefined and ez, Andres
Journal: Scientific reports (2016)
Authors: Bogert, Nicolai V and Werner, Isabella and Kornberger, Angela and Meybohm, Patrick and Moritz, Anton and Keller, Till and Stock, Ulrich A and Beiras-Fern, undefined and ez, Andres
Journal: Scientific reports (2016)
Inhibition of ABC transport proteins by oil sands process affected water
Authors: Alharbi, Hattan A and Saunders, David MV and Al-Mousa, Ahmed and Alcorn, Jane and Pereira, Alberto S and Martin, Jonathan W and Giesy, John P and Wiseman, Steve B
Journal: Aquatic Toxicology (2016): 81--88
Authors: Alharbi, Hattan A and Saunders, David MV and Al-Mousa, Ahmed and Alcorn, Jane and Pereira, Alberto S and Martin, Jonathan W and Giesy, John P and Wiseman, Steve B
Journal: Aquatic Toxicology (2016): 81--88
Rapid generation of collagen-based microtissues to study cell--matrix interactions
Authors: Brett, Marie-Elena and Crampton, Alex and ra L , undefined and Wood, David K
Journal: Technology (2016): 1--8
Authors: Brett, Marie-Elena and Crampton, Alex and ra L , undefined and Wood, David K
Journal: Technology (2016): 1--8
Toxicokinetics and toxicodynamics of chlorpyrifos is altered in embryos of Japanese medaka exposed to oil sands process-affected water: evidence for inhibition of P-glycoprotein
Authors: Alharbi, Hattan A and Alcorn, Jane and Al-Mousa, Ahmed and Giesy, John P and Wiseman, Steve B
Journal: Journal of Applied Toxicology (2016)
Authors: Alharbi, Hattan A and Alcorn, Jane and Al-Mousa, Ahmed and Giesy, John P and Wiseman, Steve B
Journal: Journal of Applied Toxicology (2016)
Flexible Endoscopic Spray Application of Respiratory Epithelial Cells as Platform Technology to Apply Cells in Tubular Organs
Authors: Thiebes, Anja Lena and Reddemann, Manuel Armin and Palmer, Johannes and Kneer, Reinhold and Jockenhoevel, Stefan and Cornelissen, Christian Gabriel
Journal: Tissue Engineering Part C: Methods (2016): 322--331
Authors: Thiebes, Anja Lena and Reddemann, Manuel Armin and Palmer, Johannes and Kneer, Reinhold and Jockenhoevel, Stefan and Cornelissen, Christian Gabriel
Journal: Tissue Engineering Part C: Methods (2016): 322--331
References
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Application notes
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
Annexin V
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
Annexin V