ATP Assay Kit ab83355 is a mix-and-read, no-wash assay with one 30 minute incubation. In the assay, glycerol is phosphorylated, with a colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) assay readout.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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Publications
- Journal of neurochemistry 168:899-9092024Bioenergetic and excitotoxic determinants of cofilactin rod formation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNguyen Mai,Long Wu,Gökhan Uruk,Ebony Mocanu,Raymond A SwansonPubMed 38299375
- Cell communication and signaling : CCS 22:512024NAT10 mediated ac4C acetylation driven mA modification via involvement of YTHDC1-LDHA/PFKM regulates glycolysis and promotes osteosarcoma.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZhongting Mei,Zhihua Shen,Jiaying Pu,Qian Liu,Guoxin Liu,Xuting He,Yang Wang,Jinrui Yue,Shiyu Ge,Tao Li,Ye Yuan,Lei YangPubMed 38233839
Key facts
- Detection method
Colorimetric/Fluorometric
- Sample types
Urine, Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate
- Assay type
Quantitative
- Assay time
1h
- Sensitivity
< 1 µM
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ATP Assay Kit ab83355 is a mix-and-read, no-wash assay with one 30 minute incubation. In the assay, glycerol is phosphorylated, with a colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) assay readout.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Key facts
- Detection method
Colorimetric/Fluorometric
- Sample types
Urine, Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate
- Assay type
Quantitative
- Assay time
1h
- Assay Platform
Microplate reader
- Sensitivity
< 1 µM
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
- Storage information
-20°C
Notes
ATP Assay Kit (Colorimetric/Fluorometric) (ab83355) use a robust, simple method; the ATP assay protocol relies on the phosphorylation of glycerol to generate a product that is easily quantified by colorimetric (ODmax = 570 nm) or fluorometric (Ex/Em = 535/587 nm) methods.
This kit can detect as low as 1 μM of ATP in various samples.
ATP assay protocol summary:
- add samples (deproteinized) and standards to wells
- add reaction mix and incubate for 30 min at room temp
- analyze with microplate reader
If you require a more sensitive product, we recommend Luminescent ATP Detection Assay Kit (Luminescent ATP Detection Assay Kit ab113849), which can detect as low as 1 pM of ATP.
**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. **How other researchers have used ATP Assay Kit ab83355** This ATP assay kit has been used in publications in a variety of sample types, including: - Human: cell culture lysates1, primary monocyte cell culture lysates2, HCT116 cell culture supernatants3 - Mouse: heart tissue4, liver5, C2C12 and L929 cell lysates6, primary thymocyte cell culture lysates7, cardiac tissue8 - Rat: primary hippocampal neuron cell culture lysates9, liver tissue10, skeletal muscle11 - Pig: kidney cell culture lysates12, heart tissue13 - C elegans tissue14 *- Chlamydomonas reinhardtii *algae15 References: 1 - Civallero M et al 2017, Na JY et al 2018, 2Gkirtzimanaki K et al 2018, 3Yang et al 2018; 4 - Singh SP et al 2018; 5 - Han SJ et al 2018; 6 - Alhindi Y et al 2019, Gregorczyk KP et al 2018; 7 - Simula L et al 2018; 8 - Litt MJ et al 2017, Samokhvalov V et al 2018; 9 - Zhao X et al 2018; 10 - Jing R et al 2018; 11 - Trinchese G et al 2018; 12 - Zou X et al 2018; 13 - Yuan F et al 2018; 14 - Pandey et al 2018; 15 - Ramanan R et al 2018
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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8 product images
Image courtesy of Mrs. Fotini MouthFunctional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
ATP levels in Pancreatic Islet.
ab83355 was used to determin ATP levels in rat pancreas islets as an ischemic marker to predict transplantation outcomes. We extracted ATP from fresh pancreas that have undergo different time of cold ischemia : 0, 2, 4, 6, 8 and 10h and in situ. ATP were extracted in Perchloric acid (PCA-2M) and grind using a Polytron. PCA were removed using potassium hydroxide (KOH – 2M) and pH was adjust around 7-8. Samples were conserved at -80°C before utilization.
Image courtesy of Maiti A K et al. PLoS One. 2018; 13(9): e0204567. doi: 10.1371/journal.pone.0204567Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
ATP assay used to study mitochondrial disfunction after C. rodentium infection in mice.
Maiti AK et al (2018) used ATP assay kit ab83355 to measure mitochondrial ATP generation in an in vitro mouse intestinal model treated with cytokines in the presence and absence of VIP (vasoactive intestinal peptide). VIP was induced by C. rodentium infection and cytokines.
Image courtesy of Maiti A K et al. Sci Rep. 2015; 5: 1543. doi: 10.1038/srep15434. Reproduced under the Creative Commons License http://creativecommons.org/licenses/by/4.0/Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
ATP levels measured in mouse colon tissue.
Maiti AK et al. (2015) used ATP assay kit TMRE-Mitochondrial Membrane Potential Assay Kit ab113852 to measure mitochondrial ATP generation in murine distal colon after C. rodentium infection.
Sanokawa-Akakura R et al., PLoS One 9(9), fig4b. doi: 10.1371/journal.pone.0108537 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
The chart shows a comparison of ATP levels of HepG2 treated with 0, 10 and 100 μM for 48 hours, DRH2O2 W1 (damage recovered cells using hydrogen peroxide with a recovery time of one week) HepG2 cells and MDA-MB-231 cells treated with 0, 10 and 100 μM of NaHS for 48 hours. Data is shown as percent of ATP levels in untreated cells. ATP levels were determined using ATP assay kit (ab83355).
Image from Shi Z et al., PLoS One 9(8), Fig 2 doi: 10.1371/journal.pone.0105675. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
MCF-7 cells are transfected with vector, osteopontin-a, osteopontin-c or osteopontin -a plus -c. Cells are plated on poly-HEMA and seeded at 4 x 105 cells per well and incubated for two days under standard culture conditions. ATP levels are measured using ATP assay kit (ab83355).
Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
Example of fluorometric ATP assay standard curve.
Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
Example of colorimetric ATP assay standard curve.
Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)
Quantitation of ATP in fish liver (2.5μl of 10 times diluted sample), fish muscle (5μl of 10 times diluted sample) and Jurkat cell lysate (5 ul) using fluorometric assay. Samples were spiked with known amounts of ATP (300pmol).
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