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| Sample Type(s) | Cells etc |
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Overview
For quantitative bioluminescent assay for ADP:ATP ratio (apoptosis) in cells and screen for modulators. The assay uses Luminescence for signal readout. Compatible sample input includes Cells etc. Typical stated assay timing is 20 min.
Key elements and design rationale
- Readout format: Luminescence supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Cells etc, which is useful when aligning matrix type with calibration and control design.
- Workflow timing: The listed assay time of 20 min helps frame batch planning, replicate handling, and plate throughput.
- Feature emphasis: Safe. Non-radioactive assay.
Additional feature notes highlight Homogeneous and convenient. “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved; Robust and amenable to HTS: Z factors of 0.5 and above are routinely observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of enzylight adp/atp ratio within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
Changes in the ADP/ATP ratio have been used to differentiate modes of cell death and viability. Increased levels of ATP and decreased levels of ADP signify proliferating cells. Conversely, decreased levels of ATP and increased levels of ADP represent apoptotic or necrotic cells where the decrease in ATP and increase in ADP is much more pronounced in necrosis versus apoptosis. BioAssay Systems’ EnzyLight™ ADP/ATP Ratio Assay Kit provides a rapid method to measure ADP and ATP levels for the screening of apoptosis, necrosis, and cell proliferation in mammalian cells. The assay involves two steps. In the first step, the working reagent lyses cells to release ATP and ADP. In the presence of luciferase, ATP immediately reacts with the Substrate D-luciferin to produce light. The light intensity is a direct measure of intracellular ATP concentration. In the second step, the ADP is converted to ATP through an enzyme reaction. This newly formed ATP then reacts with the D-luciferin as in the first step. This non-radioactive, homogeneous cell-based assay is performed in microplates. The reagent is compatible with all culture media and with all liquid handling systems for high-throughput screening applications in 96-well and 384-well plates.
Detection method
Luminescence.
Procedures and timing
Stated procedure or timing information: 20 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify enzylight adp/atp ratio in cells by Luminescence readout.
- Compare treatment or phenotype groups using matched cells handling.
- Monitor time-course or pre/post changes in cells across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
We would like to measure ADP, ATP and ADP/ATP Ratio in serum or plasma. This kit seems to be suitable for cell samples, but is it compatible with serum or plasma?
The ADP/ATP ratio assay was developed as a cell based assays and is useful to follow changes in the ADP/ATP ratio as a marker for viability in cells in the course of a drug treatment or during culture. The assay also works with serum samples but because it is not quantitative this may limit its usefulness for serum analysis. Depending on the exact nature of your experiment, our EATP-100 and EADP-100 assay kits may be the better choice.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Melatonin inhibits formation of mitochondrial permeability transition pores and improves oxidative phosphorylation of frozen-thawed ram sperm
Fang, Y., et al (2020). Melatonin inhibits formation of mitochondrial permeability transition pores and improves oxidative phosphorylation of frozen-thawed ram sperm. Frontiers in Endocrinology, 10. Assay: ATP/ADP in ram semen.
Altered energy metabolism during early optic nerve crush injury: Implications of warburg-like aerobic glycolysis in facilitating retinal ganglion cell survival
Zhu, J., et al (2020). Altered energy metabolism during early optic nerve crush injury: Implications of warburg-like aerobic glycolysis in facilitating retinal ganglion cell survival. Neuroscience Bulletin, 36(7), 761-777. Assay: ATP/ADP in mouse tissue.
RGS12 is required for the maintenance of mitochondrial function during skeletal development
Yuan, G., et al. (2020). RGS12 is required for the maintenance of mitochondrial function during skeletal development. Cell Discovery, 6(1) Assay: ATP/ADP in mouse cells.
Upregulation of DPY30 promotes cell proliferation and predicts a poor prognosis in cholangiocarcinoma
Hong, Z., et al (2020). Upregulation of DPY30 promotes cell proliferation and predicts a poor prognosis in cholangiocarcinoma. Biomedicine & Pharmacotherapy, 123, 109766. Assay: ATP/ADP in human cancer cells.
Genome reduction enhances production of polyhydroxyalkanoate and alginate oligosaccharide in pseudomonas mendocina
Fan, X., et al (2020). Genome reduction enhances production of polyhydroxyalkanoate and alginate oligosaccharide in pseudomonas mendocina. International Journal of Biological Macromolecules, 163, 2023-2031 Assay: ATP/ADP in bacteria cells.
Roles for plant mitochondrial alternative oxidase under Normoxia, hypoxia, and Reoxygenation conditions
Jayawardhane, J., et al (2020). Roles for plant mitochondrial alternative oxidase under Normoxia, hypoxia, and Reoxygenation conditions. Frontiers in Plant Science, 11. Assay: ATP/ADP in plant tissue.
Mitochondria as target for tumor management of Hemangioendothelioma
Gordillo, G. M., et al (2021). Mitochondria as target for tumor management of Hemangioendothelioma. Antioxidants & Redox Signaling, 34(2), 137-153. Assay: ATP/ADP in mouse cells.
ACE overexpression in myeloid cells increases oxidative metabolism and cellular ATP
Cao, D., et al (2019). ACE overexpression in myeloid cells increases oxidative metabolism and cellular ATP. Journal of Biological Chemistry, 295(5), 1369-1384. Assay: ATP/ADP in mouse white blood cells.
The ADP-binding kinase region of Ire1 directly contributes to its responsiveness to endoplasmic Reticulum stress
Le, Q. G., et al (2021). The ADP-binding kinase region of Ire1 directly contributes to its responsiveness to endoplasmic Reticulum stress. Scientific Reports, 11(1). Assay: ATP/ADP in yeast cells.
Transcriptional and metabolic changes associated with Phytoglobin expression during germination of Barley seeds
Zafari, S., et al (2020). Transcriptional and metabolic changes associated with Phytoglobin expression during germination of Barley seeds. International Journal of Molecular Sciences, 21(8), 2796. Assay: ATP/ADP in plant tissue.
The role of SIRT1 in avoiding AIM2-mediated antiviral defense in cervical cancer (Doctoral dissertation)
Dae-Ho So (2018). The role of SIRT1 in avoiding AIM2-mediated antiviral defense in cervical cancer (Doctoral dissertation). Assay: ADP/ATP ratio in human cells.
Metabolic reprogramming of acute lymphoblastic leukemia cells in response to glucocorticoid treatment
Dyczynski, M., Vesterlund, M., Bjorklund, A. C., Zachariadis, V., Janssen, J., Gallart-Ayala, H. & Tamm, K. P. (2018). Metabolic reprogramming of acute lymphoblastic leukemia cells in response to glucocorticoid treatment. Cell death & disease, 9(9), 846. Assay: ADP/ATP ratio in human cells.
Genome-Wide Analysis of Transcriptional Changes and Genes That Contribute to Fitness during Degradation of the Anthropogenic Pollutant Pentachlorophenol by Sphingobium chlorophenolicum
Flood, J. J., & Copley, S. D. (2018). Genome-Wide Analysis of Transcriptional Changes and Genes That Contribute to Fitness during Degradation of the Anthropogenic Pollutant Pentachlorophenol by Sphingobium chlorophenolicum. mSystems, 3(6), e00275-18. Assay: ADP/ATP ratio in S. chlorophenolicum cells.
A surfactant polymer dressing potentiates antimicrobial efficacy in biofilm disruption
Ghatak, P. D., Mathew-Steiner, S. S., Pandey, P., Roy, S., & Sen, C. K. (2018). A surfactant polymer dressing potentiates antimicrobial efficacy in biofilm disruption. Scientific reports, 8(1), 873. Assay: ADP/ATP ratio in bacterial cells.
Glycemic variability promotes both local invasion and metastatic colonization by pancreatic ductal adenocarcinoma
Jian, Z., Cheng, T., Zhang, Z., Raulefs, S., Shi, K., Steiger, K. & Bruns, P. (2018). Glycemic variability promotes both local invasion and metastatic colonization by pancreatic ductal adenocarcinoma. Cellular and Molecular Gastroenterology and Hepatology, 6(4), 429-449. Assay: ADP/ATP ratio in murine cells.
CFTR Deletion Confers Mitochondrial Dysfunction and Disrupts Lipid Homeostasis in Intestinal Epithelial Cells
Kleme, M., Sane, A., Garofalo, C., Seidman, E., Brochiero, E., Berthiaume, Y., & Levy, E. (2018). CFTR Deletion Confers Mitochondrial Dysfunction and Disrupts Lipid Homeostasis in Intestinal Epithelial Cells. Nutrients, 10(7), 836. Assay: ADP/ATP ratio in human cells.
Role of CLK3 in HIF1alpha driven cardiac hypertrophy (Doctoral dissertation, ETH Zurich)
Dominguez Sanchez, J. M. (2017). Role of CLK3 in HIF1alpha driven cardiac hypertrophy (Doctoral dissertation, ETH Zurich). Assay: ADP/ATP ratio in mouse cells.
Dominant oncogenic signal-mediated nutritional dependency affects malignant behaviours of pancreatic cancer (Doctoral dissertation, Technische Universitat Munchen)
Jian, Z. (2017). Dominant oncogenic signal-mediated nutritional dependency affects malignant behaviours of pancreatic cancer (Doctoral dissertation, Technische Universitat Munchen). Assay: ADP/ATP ratio in murine cells.
Insights into pancreatic beta cell energy metabolism using rodent beta cell models
Morten, K. J., Potter, M., Badder, L., Sivathondan, P., Dragovic, R., Neumann, A. & Lodge, T. A. (2017). Insights into pancreatic beta cell energy metabolism using rodent beta cell models. Wellcome Open Research, 2. Assay: ADP/ATP ratio in rodent cells.
Investigating IL-15 Metabolic Impact and its Mechanism of Action in Skeletal Muscle Cells (Doctoral dissertation, Universite d’Ottawa/University of Ottawa)
Nadeau, L. (2017). Investigating IL-15 Metabolic Impact and its Mechanism of Action in Skeletal Muscle Cells (Doctoral dissertation, Universite d’Ottawa/University of Ottawa). Assay: ADP/ATP ratio in rat cells.
Endocrine disruptors induce perturbations in endoplasmic reticulum and mitochondria of human pluripotent stem cell derivatives
Rajamani, U., Gross, A. R., Ocampo, C., Andres, A. M., Gottlieb, R. A., & Sareen, D. (2017). Endocrine disruptors induce perturbations in endoplasmic reticulum and mitochondria of human pluripotent stem cell derivatives. Nature communications, 8(1), 219. Assay: ADP/ATP ratio in human cells.
Novel phosphate-activated macrophages prevent ectopic calcification by increasing extracellular ATP and pyrophosphate
Villa-Bellosta, R., Hamczyk, M. R., & Andres, V. (2017). Novel phosphate-activated macrophages prevent ectopic calcification by increasing extracellular ATP and pyrophosphate. PloS one, 12(3), e0174998. Assay: ADP/ATP ratio in mice cells.
Capsaicin triggers autophagic cell survival which drives epithelial mesenchymal transition and chemoresistance in bladder cancer cells in an Hedgehog-dependent manner
Amantini, C., Morelli, M. B., Nabissi, M., Cardinali, C., Santoni, M., Gismondi, A., & Santoni, G. (2016). Capsaicin triggers autophagic cell survival which drives epithelial mesenchymal transition and chemoresistance in bladder cancer cells in an Hedgehog-dependent manner. Oncotarget, 7(31), 50180. Assay: ADP/ATP ratio in human cells.
Lactate’s effect on human neuroblastoma cell bioenergetic fluxes
Lezi, E., & Swerdlow, R. H. (2016). Lactate’s effect on human neuroblastoma cell bioenergetic fluxes. Biochemical pharmacology, 99, 88-100. Assay: ADP/ATP ratio in human cells.
MiR-186 inhibited aerobic glycolysis in gastric cancer via HIF-1alpha regulation
Liu, L., Wang, Y., Bai, R., Yang, K., & Tian, Z. (2016). MiR-186 inhibited aerobic glycolysis in gastric cancer via HIF-1alpha regulation. Oncogenesis, 5(5), e224. Assay: ADP/ATP ratio in human cells.
Interplay of mtDNA, metabolism and microbiota in the pathogenesis of AIBD (Doctoral dissertation, Universitat zu Lubeck)
Schilf, P. (2016). Interplay of mtDNA, metabolism and microbiota in the pathogenesis of AIBD (Doctoral dissertation, Universitat zu Lubeck). Assay: ADP/ATP ratio in mice cells.
Prognostic significance and function of the vacuolar H+-ATPase subunit V1E1 in esophageal squamous cell carcinoma
Son, S. W., Kim, S. H., Moon, E. Y., Kim, D. H., Pyo, S., & Um, S. H. (2016). Prognostic significance and function of the vacuolar H+-ATPase subunit V1E1 in esophageal squamous cell carcinoma. Oncotarget, 7(31), 49334. Assay: ADP/ATP ratio in human cells.
mTOR hyperactivation by ablation of tuberous sclerosis complex 2 in the mouse heart induces cardiac dysfunction with the increased number of small mitochondria mediated through the down-regulation of autophagy
Taneike, M., Nishida, K., Omiya, S., Zarrinpashneh, E., Misaka, T., Kitazume-Taneike, R. & Shah, A. M. (2016). mTOR hyperactivation by ablation of tuberous sclerosis complex 2 in the mouse heart induces cardiac dysfunction with the increased number of small mitochondria mediated through the down-regulation of autophagy. PLoS One, 11(3), e0152628. Assay: ADP/ATP ratio in mice tissues.
Apyrase therapy for bleeding conditions
Chen, R. & Jeong, SS (2013). Apyrase therapy for bleeding conditions. WO/2011/088231. Assay: ADP/ATP ratio in human cell.
Dietary protein restriction induces steatohepatitis and alters leptin/signal transducers and activators of transcription 3 signaling in lactating rats
Kwon DH et al (2012).Dietary protein restriction induces steatohepatitis and alters leptin/signal transducers and activators of transcription 3 signaling in lactating rats. J Nutr Biochem 23(7):791-9. Assay: ADP/ATP ratio in human cell.
Pyrase Therapy For Bleeding Conditions
Chen, R. and Jeong, S. (2011). Pyrase Therapy For Bleeding Conditions. WO/2011/088231. Assay: ADP/ATP ratio in rat bronchoalveolar fluid.
Glabridin protects osteoblastic MC3T3-E1 cells against antimycin A induced cytotoxicity
Choi EM (2011). Glabridin protects osteoblastic MC3T3-E1 cells against antimycin A induced cytotoxicity. Chem Biol Interact. 193(1):71-8. Assay: ADP/ATP ratio in mouse cell line.
Luteolin protects osteoblastic MC3T3-E1 cells from antimycin A-induced cytotoxicity through the improved mitochondrial function and activation of PI3K/Akt/CREB
Choi EM (2011). Luteolin protects osteoblastic MC3T3-E1 cells from antimycin A-induced cytotoxicity through the improved mitochondrial function and activation of PI3K/Akt/CREB. Toxicol In Vitro. 25(8):1671-9. Assay: ADP/ATP ratio in mouse cell line.
Luteolin protects osteoblastic MC3T3-E1 cells from antimycin A-induced cytotoxicity through the improved mitochondrial function and activation of PI3K/Akt/CREB
Choi EM (2011). Luteolin protects osteoblastic MC3T3-E1 cells from antimycin A-induced cytotoxicity through the improved mitochondrial function and activation of PI3K/Akt/CREB. Toxicol In Vitro. Assay: ADP/ATP ratio in rat bronchoalveolar fluid.
Methods for treatment of metabolic disorders using epimetabolic shifters, multidimensional intracellular molecules, or environmental influencers
Narain, NR, Mccook, JP, Sarangarajan, R. et al (2011). Methods for treatment of metabolic disorders using epimetabolic shifters, multidimensional intracellular molecules, or environmental influencers. US Patent Appl. 20110020312. Assay: ADP/ATP ratio in human cell, environmental.
Oxygen-inducible glutamate oxaloacetate transaminase as protective switch transforming neurotoxic glutamate to metabolic fuel during acute ischemic stroke
Rink, C et al (2011). Oxygen-inducible glutamate oxaloacetate transaminase as protective switch transforming neurotoxic glutamate to metabolic fuel during acute ischemic stroke. Antioxid Redox Signal 14(10):1777-85. Assay: ADP/ATP ratio in mouse cell line.
Methods for treatment of a sarcoma using an epimetabolic shifter (Coenzyme q10)
Sarangarajan, R. (2011). Methods for treatment of a sarcoma using an epimetabolic shifter (Coenzyme q10). US 2011/0064747. Assay: ADP/ATP ratio in human cell.
Conditional HIF-1alpha expression produces a reversible cardiomyopathy
Bekeredjian R, Suhr ST et al (2010). Conditional HIF-1alpha expression produces a reversible cardiomyopathy. PLoS One 5(7):e11693. Assay: ADP/ATP ratio in human cell line.
Conditional HIF-1alpha expression produces a reversible cardiomyopathy
Bekeredjian R. et al. (2010). Conditional HIF-1alpha expression produces a reversible cardiomyopathy. PLoS One 5(7):e11693. Assay: ADP/ATP ratio in mouse heart lysate.
Efficient phagocytosis requires triacylglycerol hydrolysis by adipose triglyceride lipase
Chandak PG et al (2010). Efficient phagocytosis requires triacylglycerol hydrolysis by adipose triglyceride lipase. J Biol Chem. 285(26):20192-201. Assay: ADP/ATP ratio in rat total ATP/ADP in mitochondria.
Efficient phagocytosis requires triacylglycerol hydrolysis by adipose triglyceride lipase
Chandak PG, Suhr ST, et al. (2010). Efficient phagocytosis requires triacylglycerol hydrolysis by adipose triglyceride lipase. J Biol Chem. 285(26):20192-201. Assay: ADP/ATP ratio in mouse cells.
Inhibitory effects of adenine nucleotides on brain mitochondrial permeability transition
Saito A, Castilho RF (2010). Inhibitory effects of adenine nucleotides on brain mitochondrial permeability transition. Neurochem Res. 35(11):1667-74. Assay: ADP/ATP ratio in mouse heart lysate.
Mitochondrial rejuvenation after induced pluripotency
Suhr ST et al (2010). Mitochondrial rejuvenation after induced pluripotency. PLoS One 5(11):e14095. Assay: ADP/ATP ratio in rat brain tissue.
Mitochondrial rejuvenation after induced pluripotency
Suhr ST, et al. (2010). Mitochondrial rejuvenation after induced pluripotency. PLoS One 5(11):e14095. Assay: ADP/ATP ratio in human cell line.
Antioxidant and oncogene rescue of metabolic defects caused by loss of matrix attachment
Schafer ZT, et al (2009). Antioxidant and oncogene rescue of metabolic defects caused by loss of matrix attachment. Nature. 461(7260):109-113. Assay: ADP/ATP ratio in human cell.
Apyrase treatment prevents ischemia-reperfusion injury in rat lung isografts
Sugimoto S et al (2009). Apyrase treatment prevents ischemia-reperfusion injury in rat lung isografts. J Thorac Cardiovasc Surg.138(3):752-9. Assay: ADP/ATP ratio in mouse cells.
Apyrase treatment prevents ischemia-reperfusion injury in rat lung isografts
Sugimoto S, et al. (2009). Apyrase treatment prevents ischemia-reperfusion injury in rat lung isografts. J Thorac Cardiovasc Surg.138(3):752-9. Assay: ADP/ATP ratio in rat lung tissue.
