Luminescent ATP Detection Assay Kit (ab113849) is used to measure the level of ATP within the cell.
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Publications
- The FEBS journal 291:2221-22412024Icariin ameliorates glycolytic dysfunction in Alzheimer's disease models by activating the Wnt/β-catenin signaling pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJu Liu,Ai-Hong Wei,Ting-Ting Liu,Xin-Hao Ji,Ying Zhang,Fei Yan,Mei-Xiang Chen,Jin-Bo Hu,Shao-Yu Zhou,Jing-Shan Shi,Hai Jin,Feng JinPubMed 38400523
- The Journal of nutritional biochemistry 124:1095352023Raspberry polyphenols target molecular pathways of heart failure.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRami S Najjar,Ranjan K Roy,Javier E Stern,Rafaela G FeresinPubMed 37984734
Key facts
- Detection method
Luminescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Quantitative
- Assay time
30m
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Luminescent ATP Detection Assay Kit (ab113849) is used to measure the level of ATP within the cell.
Key facts
- Detection method
Luminescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Quantitative
- Assay time
30m
- Assay Platform
Microplate reader
Storage
- Shipped at conditions
Blue Ice
- Appropriate long-term storage conditions
+4°C
- Storage information
+4°C
Notes
Luminescent ATP Detection Assay Kit (ab113849) is used to measure the level of ATP within the cell. The luminescent ATP assay protocol involves lysis of the cell sample, addition of luciferase enzyme and luciferin, and measurement of the emitted light using a tube or microplate-based luminometer.
This kit irreversibly inactivates ATP degrading enzymes (ATPases) during the lysis step, ensuring that the luminescent signal obtained truly corresponds to the endogenous levels of ATP.
Luminescent ATP assay protocol summary:
- add ATP standard into standard wells and media into control wells in same plate containing cells to be analyzed
- add detergent solution and incubate for 5 min to lyse cells and stabilize ATP
- add substrate solution and incubate for 5 min
- store plate in dark for 10 min
- analyze on luminescence plate reader
Special Handling Instructions for the ATP Detection Assay Kit
ATP can be found in cells and microbiota on many surfaces. To prevent unintended background, it is recommended to clean bench surfaces and all pipettes to be used during the experiment with 10% bleach. Use of gloves first cleaned by either using 70% ethanol or by changing them frequently is recommended. Use tips and containers that are clean and sterile, such as ATP and nuclease-free consumables. Do not leave reagents or the plate opened while working or during assay incubation
Total levels of cellular ATP can be used to assess cell viability, cell proliferation and cytotoxicity of a wide range of compounds and biological response modifiers. We also offer a very popular alternative colorimetric/fluorometric based on the phosphorylation of glycerol. **Related assays** Review the to learn about kits to perform a , and . Review the to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
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It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
- Activity summary
ATP or adenosine triphosphate acts mechanically as the key energy carrier in cellular processes. It has a molecular mass of approximately 507 Da. ATP is present throughout prokaryotic and eukaryotic cells particularly in the cytoplasm mitochondria and chloroplasts. Known also as the "molecular unit of currency" ATP transfers energy for all biochemical reactions and cellular activities. ATP functions in various cellular activities including active transport across membranes signal transduction and muscle contraction.
- Biological function summary
ATP provides the energy necessary for most cellular functions. It is part of the ATP synthesis complex also known as ATP synthase localized in the mitochondrial inner membrane (in eukaryotes) and the plasma membrane (in prokaryotes). This complex facilitates ATP production through oxidative phosphorylation. ATP also plays a significant role in cellular metabolism and energy transfer. Many enzymes use ATP to catalyze reactions converting ATP to ADP and inorganic phosphate releasing energy to power cellular activities.
- Pathways
ATP is integral to both glycolysis and the citric acid cycle. In glycolysis ATP participates in substrate-level phosphorylation transferring energy from glucose breakdown steps. The citric acid cycle further processes metabolic intermediates generating high-energy electron carriers that feed the electron transport chain. This chain leads to ATP production via oxidative phosphorylation. ATP interacts with enzymes like hexokinase and phosphofructokinase in glycolysis and other enzymes in the electron transport chain like cytochrome c oxidase.
- Associated diseases and disorders
ATP's role is linked to various conditions such as mitochondrial diseases and cardiac ischemia. In mitochondrial diseases mutations in mitochondrial DNA affect ATP production leading to energy deficits in cells. These disorders involve proteins like NADH dehydrogenase a component of the mitochondrial respiratory chain. In cardiac ischemia reduced oxygen supply impairs ATP synthesis affecting heart muscle contraction. Ischemia involves proteins such as creatine kinase which buffers ATP levels to sustain cellular energy during reduced blood flow.
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5 product images
D?ini?, Tamara and Norbert A Dencher., Oxidative medicine and cellular longevity?vol. 2018 7567959., Fig 6, doi:10.1155/2018/7567959Functional Studies - Luminescent ATP Detection Assay Kit (ab113849)
Total cellular ATP concentration. ATP in SH-SY5Y cells cultivated at 21% and 5% O2 24 h after treatment with A? peptide and/or 18 h X-ray irradiation, normalized to cell count, and compared to respective controls. ATP concentration was about 1.3- to 1.8-fold higher at all conditions in cells cultivated at 5% O2 compared to 21% O2. Combination of A? peptide treatment and irradiation resulted in a significantly increased (~1.5-fold) ATP concentration at 5% O2 compared to the control. Samples were measured at least in duplicates (n = 2-4) in three independent experiments (N = 3). Mean ± SEM analyzed by two-way ANOVA with Tukey's multiple comparison test with p < 0 05 considered as significant. (∗∗p < 0 01).
Cellular Activity - Luminescent ATP Detection Assay Kit (ab113849)
Example of ATP standard curve using an opaque white plate.
The ATP standard curve was prepared as described in the protocol. Background-subtracted data values (mean +/- SD) are graphed.
Functional Studies - Luminescent ATP Detection Assay Kit (ab113849)
ab113849 ATP detection kit cytotoxicity data. 25000 HepG2 cells were seeded into each well, allowed to adhere and treated for 4 hours with 25µM rotenone and vehicle control (DMSO) in glucose based complete media. After treatment, cells were lysed, exposed to the ATP substrate solution and signal was measured on a luminescent counter. Mean and standard deviation is plotted for 3 replicates from each condition. Rotenone induces cytotoxicity in HepG2 cells.
Cellular Activity - Luminescent ATP Detection Assay Kit (ab113849)
Simultaneous quantification of mitochondrial respiration and glycolytic flux.
Cellular Energy Flux for HepG2 cells (seeded at 65,000 per well), treated with a combination of drug compounds modulating the ETC (Antimycin A [1 μM] and FCCP [2.5 μM]), shown as a percentage relative to untreated control cells. Comparative measurements were taken with Extracellular Oxygen Consumption Assay (Extracellular Oxygen Consumption Assay ab197243) (white column) and Glycolysis Assay [Extracellular acidification] (Glycolysis Assay [Extracellular acidification] ab197244) (black column) show the shift between mitochondrial respiration and glycolysis and the cellular control of energy (ATP; measured 1h post-treatment using Luminescent ATP Detection Assay kit (ab113849) (striped column)).
This image is courtesy of a customer review submitted by Heiko Lemcke.Functional Studies - Luminescent ATP Detection Assay Kit (ab113849)
Analysis of the release of ATP by connexin hemichannels in stem cells using ATP luminescence kit (ab113849).
Cells were cultured in HBSS to induce hemichannel opening. Calcium and GAP-inhibitor were used to trigger hemichannel closure.
After two hours the supernatant was collected and ATP was measured according to the protocol (detergent was also applied).Calcium treatment and inhibition by GAP decreased ATP concentration, compared to HBSS control. Graph shows data of three independent experiments.
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