Calculator
Cell Meter™ Live Cell ATP Assay Kit
Example protocol
AT A GLANCE
Prepare cells in a growth medium.
Incubate the cells with the ATP Red™ working solution at 37°C for 15 to 30 minutes.
Remove the ATP Red™ working solution.
Analyze with a fluorescence microscope using a Cy3/TRITC filter set.
CELL PREPARATION
Plate cells overnight in a growth medium. For a 96-well plate, use 10,000 to 40,000 cells/well/90 μL. For a 384-well plate, use 2,500 to 10,000 cells/well/20 μL.
Centrifuge the cells from the culture medium. Suspend the cell pellets in culture medium, preparing 50,000-100,000 cells/well/90 µL for a 96-well poly-D lysine plate or 10,000-25,000 cells/well/20 µL for a 384-well poly-D lysine plate. Before starting your experiment, centrifuge the plate at 800 rpm for 2 minutes with the brake off.
Note: Each cell line should be individually assessed to determine the optimal cell density.
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 50 µL of DMSO (Component C) to the vial containing ATP Red™ (Component A) to prepare a 200X stock solution.
Note: 50 µL of the 200X ATP Red™ stock solution is sufficient for one 96-well plate. Any unused ATP Red™ stock solution can be aliquoted and stored at ≤ -20°C, protected from light. Avoid freeze/thaw cycles.
PREPARATION OF WORKING SOLUTION
Add 5 µL of the 200X stock solution (Component A) to 1 mL of cell culture medium and mix thoroughly.
Note: The ATP Red™ probe is suitable for use with the cell culture media of most cell lines we have tested. Staining conditions can be adjusted based on the specific cell type.
SAMPLE EXPERIMENTAL PROTOCOL
Prepare cells in a growth medium.
Add an equal volume of ATP Red™ working solution to each well in the cell plate. Use 100 µL per well for a 96-well plate or 25 µL per well for a 384-well plate.
Note: The ideal concentration of ATP Red™ depends on the specific application.
Incubate the cells at 37°C for 15-30 minutes, protected from light.
Remove the working solution from each well. Then, wash the cells twice with Assay Buffer (Component B) or any other buffer of your choice.
Use a fluorescence microscope with a Cy3/TRITC filter set to observe the fluorescence signal in the cells
Citations
Authors: Abdelwahid, E and Chen, W and Ascoli, C and Jacobson, JR and Singla, S
Journal: American Journal of Respiratory and Critical Care Medicine (2025): A4626--A4626
Authors: Jeon, Byeong Bae and Shin, Jung Un and Shin, Dong Wook and Cho, Dong-Ha
Journal: Journal of the Society of Cosmetic Scientists of Korea (2025): 11--21
Authors: Yan, Feihong and Li, Ruiyuan and Liu, Jiaxin and Yang, Lulu and Liu, Helin and Zhu, Shengcang and Zhang, Yuhui and Wang, Lijun and Huang, Lu and Wang, Yu and others,
Journal: Advanced Composites and Hybrid Materials (2025): 173
References
Authors: Ishii, Masaaki and Beeson, Gyda and Beeson, Craig and Rohrer, Bärbel
Journal: Frontiers in immunology (2021): 628062
Authors: Richani, Dulama and Dunning, Kylie R and Thompson, Jeremy G and Gilchrist, Robert B
Journal: Human reproduction update (2021): 27-47
Authors: Guo, Yusi and Chi, Xiaopei and Wang, Yifan and Heng, Boon Chin and Wei, Yan and Zhang, Xuehui and Zhao, Han and Yin, Ying and Deng, Xuliang
Journal: Stem cell research & therapy (2020): 245
Authors: Chen, Kun-Lin and Wang, Hui-Li and Jiang, Lin-Zheng and Qian, Yong and Yang, Cai-Xia and Chang, Wei-Wei and Zhong, Ji-Feng and Xing, Guang-Dong
Journal: In vitro cellular & developmental biology. Animal (2020): 322-331
Authors: Ren, Tian-Bing and Wen, Si-Yu and Wang, Lu and Lu, Peng and Xiong, Bin and Yuan, Lin and Zhang, Xiao-Bing
Journal: Analytical chemistry (2020): 4681-4688
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