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| Sample Type(s) | Cells etc |
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Overview
For rapid, quantitative, bioluminescent determination of ATP and evaluation of drug effects on ATP metabolism. The assay uses Luminescence for signal readout. Compatible sample input includes Cells etc. Typical stated assay timing is 10 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.
- Analytical range context: The supplied specifications include a stated detection limit of 0.02 µM for interpreting low-signal samples.
- Feature emphasis: Safe. Non-radioactive assay.
Additional feature notes highlight Sensitive and accurate. As low as 0.02 µM ATP or a single cell can be quantified; Homogeneous and convenient. “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of enzylight atp 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
Adenosine 5’-triphosphate (ATP) is the chemical energy for cellular metabolism and is often referred to as “energy currency” of the cell. ATP is produced only in living cells during photosynthesis and cellular respiration and consumed in cellular processes including biosynthetic reactions, motility and cell division. It is a key indicator of cellular activity and has been utilized as a measure of cell viability and cytotoxicity in research and drug discovery. BioAssay Systems’ EnzyLight™ ATP Assay Kit provides a rapid method to measure intracellular ATP. The single working reagent lyses cells to release ATP, which, in the presence of luciferase, immediately reacts with the Substrate D-luciferin to produce light. The light intensity is a direct measure of intracellular ATP concentration. This non-radioactive, homogeneous cell-based assay is performed in microplates. The reagent is compatible with all liquid handling systems for high-throughput screening applications in 96-well and 384-well plates.
Detection method
Luminescence.
Detection limit and analytical sensitivity
Reported detection limit: 0.02 µM.
Procedures and timing
Stated procedure or timing information: 10 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 atp 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.
Can this kit be used to adhesive cells?
This kit is suitable for adherent cells. The assay buffer contains 0.5 % Triton X-100 that will efficiently lyse the cells.
Is the lysis buffer in the kit suitable for Bradford protein assay?
The final Triton X-100 concentration in the lysate will be around 0.25 % which would interfere with the Bradford assay. Therefore, I would recommend diluting the lysate further. The commonly used BCA assay is not compatible, because of the high amount of mercaptoethanol in the assay buffer (70 mM).
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.
Anti-cancer strategy targeting the energy metabolism of tumor cells surviving a low-nutrient acidic microenvironment
Maeda, Y. et al (2020). Anti-cancer strategy targeting the energy metabolism of tumor cells surviving a low-nutrient acidic microenvironment. Molecular Metabolism, 42. Assay: ATP in human lung cancer cell.
Tunicamycin-Induced Endoplasmic Reticulum Stress Mediates Mitochondrial Dysfunction in Human Adipocytes
Jackisch, L. et al (2020). Tunicamycin-Induced Endoplasmic Reticulum Stress Mediates Mitochondrial Dysfunction in Human Adipocytes. The Journal of clinical endocrinology and metabolism, 105(9), dgaa258. Assay: ATP in human adipocytes.
IF1 connects obesity and insulin resistance through mitochondrial reprogramming in association with ANT2
Wang, Y. et al (2020). IF1 connects obesity and insulin resistance through mitochondrial reprogramming in association with ANT2. Assay: ATP in mouse muscle tissue.
Antimicrobial Peptide against Mycobacterium Tuberculosis That Activates Autophagy Is an Effective Treatment for Tuberculosis
Coyotl, E. et al (2020). Antimicrobial Peptide against Mycobacterium Tuberculosis That Activates Autophagy Is an Effective Treatment for Tuberculosis. Pharmaceutics, 12(11). Assay: ATP in human HEK293T cells.
Bcl-2 Proteins Regulate Mitophagy in Lipopolysaccharide-Induced Acute Lung Injury via PINK1/Parkin Signaling Pathway
Zhang, Z. et al (2020). Bcl-2 Proteins Regulate Mitophagy in Lipopolysaccharide-Induced Acute Lung Injury via PINK1/Parkin Signaling Pathway. Oxidative Medicine and Cellular Longevity, 2020. Assay: ATP in human A549 Cell.
Regulation of microbiota-GLP1 axis by Sennoside A (SA) in diet-induced obese mice
Le, Jiamei, et al (2019). Regulation of microbiota-GLP1 axis by Sennoside A (SA) in diet-induced obese mice. Acta Pharmaceutica Sinica B. Assay: ATP in mice tissues.
Cytotoxicity of Malondialdehyde and Cytoprotective Effects of Taurine via Oxidative Stress and PGC-1alpha Signal Pathway in C2C12 Cells
Cai, J-G., et al (2018). Cytotoxicity of Malondialdehyde and Cytoprotective Effects of Taurine via Oxidative Stress and PGC-1alpha Signal Pathway in C2C12 Cells. Molecular Biology 52.4: 532-542. Assay: ATP in mouse cells.
Effects of red blood cell supernatants on hypoxia/reoxygenation injury in H9C2 cells
Fan, Fengyan, et al (2018). Effects of red blood cell supernatants on hypoxia/reoxygenation injury in H9C2 cells. Intl.J. Clin. Exp. Med.11.4: 3612-3619. Assay: ATP in rat H9C2 cells.
Inhibition of SNAT5 induces incretin-responsive state from incretin-unresponsive state in pancreatic beta-cells: study of beta-cell spheroid clusters as a model
Hashim, Mahira, et al (2018). Inhibition of SNAT5 induces incretin-responsive state from incretin-unresponsive state in pancreatic beta-cells: study of beta-cell spheroid clusters as a model. Diabetes 67.9: 1795-1806. Assay: ATP in mice pancreatic beta-cells.
Mitochondrial protein turnover is critical for granulosa cell proliferation and differentiation in antral follicles
Hoque, SA Masudul, et al (2018). Mitochondrial protein turnover is critical for granulosa cell proliferation and differentiation in antral follicles. Journal of the Endocrine Society 3.2: 324-339. Assay: ATP in mouse granulosa cell.
Increasing oxygen availability for improving poly (3-hydroxybutyrate) production by Halomonas
Ouyang, Pengfei, et al (2018). Increasing oxygen availability for improving poly (3-hydroxybutyrate) production by Halomonas. Metabolic engineering 45: 20-31. Assay: ATP in Halomonas bluephagenesis cells.
APP promotes osteoblast survival and bone formation by regulating mitochondrial function and preventing oxidative stress
Pan, Jin-Xiu, et al (2018). APP promotes osteoblast survival and bone formation by regulating mitochondrial function and preventing oxidative stress. Cell death & disease 9.11: 1077. Assay: ATP in mice cells.
Mitochondria-mediated disturbance of fatty acid metabolism in proximal tubule epithelial cells leads to renal interstitial fibrosis
Shen, W., et al (2018). Mitochondria-mediated disturbance of fatty acid metabolism in proximal tubule epithelial cells leads to renal interstitial fibrosis. Eur Rev Med Pharmacol Sci 22.3: 810-819. Assay: ATP in mice serum.
Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice
Sun, Yongning, et al (2018). Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice. Nutrition & diabetes 8.1: 53. Assay: ATP in mice tissues.
Propofol inhibits parthanatos via ROS-ER-calcium-mitochondria signal pathway in vivo and vitro
Zhong, Hanhui, et al (2018). Propofol inhibits parthanatos via ROS-ER-calcium-mitochondria signal pathway in vivo and vitro. Cell death & disease 9.10: 932. Assay: ATP in mice cells.
Protective effect of alpha-lipoic acid against antimycin A cytotoxicity in MC3T3-E1 osteoblastic cells
Lin, Zou, et al (2017). Protective effect of alpha-lipoic acid against antimycin A cytotoxicity in MC3T3-E1 osteoblastic cells. Cell Stress and Chaperones 22.1: 5-13. Assay: ATP in mouse cells.
Glutamate oxaloacetate transaminase enables anaplerotic refilling of TCA cycle intermediates in stroke-affected brain
Rink, Cameron, et al (2017). Glutamate oxaloacetate transaminase enables anaplerotic refilling of TCA cycle intermediates in stroke-affected brain. The FASEB Journal 31.4: 1709-1718. Assay: ATP in mice brain tissues.
Multiplicity of acquired cross-resistance in paclitaxel-resistant cancer cells is associated with feedback control of TUBB3 via FOXO3a-mediated ABCB1 regulation
Aldonza, Mark Borris D., et al (2016). Multiplicity of acquired cross-resistance in paclitaxel-resistant cancer cells is associated with feedback control of TUBB3 via FOXO3a-mediated ABCB1 regulation. Oncotarget 7.23: 34395. Assay: ATP in human cells.
Nuclear ULK1 promotes cell death in response to oxidative stress through PARP1
Joshi, A., et al (2016). Nuclear ULK1 promotes cell death in response to oxidative stress through PARP1. Cell death and differentiation 23.2: 216. Assay: ATP in murine cells.
Luteolin alleviates methylglyoxal-induced cytotoxicity in osteoblastic MC3T3-E1 cells
Suh, Kwang Sik, Suk Chon, and Eun Mi Choi (2016). Luteolin alleviates methylglyoxal-induced cytotoxicity in osteoblastic MC3T3-E1 cells. Cytotechnology 68.6: 2539-2552. Assay: ATP in mouse cells.
Triacylglycerol accumulation activates the mitochondrial apoptosis pathway in macrophages
Aflaki E, et al (2011). Triacylglycerol accumulation activates the mitochondrial apoptosis pathway in macrophages. J Biol Chem. 286(9):7418-28. Assay: ATP in mouse macrophage.
Protective effect of apocynin on antimycin A-induced cell damage in osteoblastic MC3T3-E1 cells
Choi EM, Lee YS (2011). Protective effect of apocynin on antimycin A-induced cell damage in osteoblastic MC3T3-E1 cells. J Appl Toxicol. 32(9):714-21. Assay: ATP in mouse cell.
Analysis of the capability fo ultra-highly diluted glucose to increase glucose uptake in arsenite-streesed bacteria Escherichia coli
Khuda-Buksh, AR. et al. (2011). Analysis of the capability fo ultra-highly diluted glucose to increase glucose uptake in arsenite-streesed bacteria Escherichia coli. J, Chin. Integrative Medicine 9(8): 901-912. Assay: ATP in bacteria cell lysate.
Analysis of the capability fo ultra-highly diluted glucose to increase glucose uptake in arsenite-streesed bacteria Escherichia coli
Khuda-Buksh, AR. et al. (2011). Analysis of the capability fo ultra-highly diluted glucose to increase glucose uptake in arsenite-streesed bacteria Escherichia coli. J, Chin. Integrative Medicine 9(8): 901-912. Assay: ATP in bacteria.
Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme q10). US Patent Appl. 20110027247
Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme q10). US Patent Appl. 20110027247. Assay: ATP in mouse cell.
Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme q10)
Narain, NR, McCook, JP. (2011). Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme q10). US Patent Appl. 20110027247. Assay: ATP in human cell.
P2X7 receptors mediate deleterious renal epithelial-fibroblast cross talk
Ponnusamy M, et al (2011). P2X7 receptors mediate deleterious renal epithelial-fibroblast cross talk. Am J Physiol Renal Physiol. 300(1):F62-70. Assay: ATP in rat fibroblast.
Role of purinergic signaling pathways in V-ATPase recruitment to apical membrane of acidifying epididymal clear cells
Belleannee C, et al (2010). Role of purinergic signaling pathways in V-ATPase recruitment to apical membrane of acidifying epididymal clear cells. Am J Physiol Cell Physiol. 298(4):C817-30. Assay: ATP in rat tube.
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: ATP in mouse cells.
Liposome-delivered ATP effectively protects the retina against ischemia-reperfusion injury
Dvoriantchikova G, et al (2010). Liposome-delivered ATP effectively protects the retina against ischemia-reperfusion injury. Mol Vis.16:2882-90. Assay: ATP in mouse retina.
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: ATP in rat lung tissue.
Oxidative stress caused by pyocyanin impairs CFTR Cl(-) transport in human bronchial epithelial cells
Schwarzer C, et al (2008). Oxidative stress caused by pyocyanin impairs CFTR Cl(-) transport in human bronchial epithelial cells. Free Radic Biol Med. 45(12):1653-62. Assay: ATP in human cell.
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