Description
Key facts
Detection method:Luminescent,
Sample types:Suspension cells, Adherent cells,
Assay type:Quantitative,
Assay time:30m,
Assay Platform:Microplate reader
Product details:
How the assay works
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
Related and recommended products
We also offer a very popular alternative colorimetric/fluorometric ATP assay kit
ab83355
based on the phosphorylation of glycerol.
How other researchers are using
Luminescent ATP Detection Assay Kit has been used in a variety of sample type including:
HeLa cells
Mouse microglia cells
Rat ventricular cardiomyoblasts
References:
1-Sittewelle M et al. 2023
2-Sabogal-Guбqueta A et al. 2023
3-Gurler B et al. 2023
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.
