ATP Assay Kit (Colorimetric/Fluorometric)
4
(5 Reviews)
|
(517 Publications)
- BIOVISION® assay kit
- Cited in over 400 publications
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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AbReview
Functional 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 Mrs. Fotini Mouth
- FuncS
PubMed
Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (AB83355)
ATP levels measured in mouse colon tissue.
Maiti AK et al. (2015) used ATP assay kit ab113852 to measure mitochondrial ATP generation in murine distal colon after C. rodentium infection.
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/
- FuncS
PubMed
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).
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/
- FuncS
PubMed
Functional 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. PLoS One. 2018; 13(9): e0204567. doi: 10.1371/journal.pone.0204567
- FuncS
Supplier Data
Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (AB83355)
Example of colorimetric ATP assay standard curve.
- FuncS
PubMed
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).
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/
- FuncS
Supplier Data
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).
- FuncS
Supplier Data
Functional Studies - ATP Assay Kit (Colorimetric/Fluorometric) (AB83355)
Example of fluorometric ATP assay standard curve.
- Biochemical assay
Lab
Biochemical assay - ATP Assay Kit (Colorimetric/Fluorometric) (AB83355)
Diagram showing the principles of the ATP assay method.
- Schematic Diagram
Supplier Data
Schematic Diagram - ATP Assay Kit (Colorimetric/Fluorometric) (AB83355)
Representative image of ATP Assay Kit (Colorimetric/Fluorometric) ab83355
Components shown from left to right :
- OxiRed Probe
- Converter Mix B
- ATP IV
- Developer Mix N
- Assay Buffer 23
Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the kit.
Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit.
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Product details
ATP Assay Kit (Colorimetric/Fluorometric) (ab83355) use a robust, simple ATP assay method. This kit can detect as low as 1 μM of ATP in various samples
How the assay works
ATP assay kit ab83355 uses the standard established assay protocol for fluorometric and colorimetric ATP determination kits:
- glycerol is phosphorylated by glycerol kinase in an ATP dependent reaction to produce glycerol 3-phosphate
- glycerol 3-phosphate is then acted on by glycerol phosphate oxidase to produce glycerone phosphate and hydrogen peroxide
- hydrogen peroxide is then processed by a peroxidase causing a reaction with a probe to generate red color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm).
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
Related ATP assay products
If you require a more sensitive product, we recommend Luminescent ATP Detection Assay Kit (ab113849), which can detect as low as 1 pM of ATP.
How other researchers are using 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
Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K354 ATP Colorimetric/Fluorometric Assay Kit. K354-100 is the same size as the 100 test size of ab83355.
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Properties and storage information
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Storage information
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Product protocols
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This is a protocol summary; please use the downloadable protocol booklet for full instructions.
ATP assay protocol
Reagent prep
Reconstitute ATP IV in water to 10mM. Reconstitute Converter Mix B and Developer Mix N in Assay Buffer 23. Other assay components are ready-to-use.
Note – DMSO tends to be solid when stored at -20°C, even when left at room temperature; melt DMSO solution (OxiRed Probe) in a 37°C bath for 1 – 5 min before use.
Sample prep
Homogenize cells by pipetting (and tissue with a Dounce homogenizer) in Assay Buffer 23. Insoluble material is removed by centrifuge, and the supernatant is used for the assay.
Use Red Blood Cell Lysis Buffer (ab204733) for red blood cells.
Cell and tissue samples may contain enzymes that interfere with the assay. Remove these enzymes from sample and deproteinize samples with TCA kit ab204708, or use PCA/KOH deproteinization.
Use heparin when collecting plasma or serum. EDTA and other chelators should be avoided. Deproteinize fluid samples with 10 kD spin columns (ab93349).
Notes:
-highly metabolically active tissues, such as muscle, should be prepared directly in PCA.
- do not extend incubation times of 5 min when using TCA. Longer incubation times may result in loss of ATP.
- fresh samples are preferable, if this is not possible then snap freeze in liquid nitrogen upon extraction and store at -80°C.
ATP assay procedure
Notes:
- detection sensitivity of fluorometric assay is 10-100 fold higher than colorimetric assay.
- glycerol phosphate present in cell or tissue extracts can generate background in this assay. If you suspect your samples contain glycerol phosphate, set up Sample Background Controls.
- OxiRed Probe is very sensitive to exposure to air and light and hence should be kept tightly sealed, and should be added to the reaction mix right before the assay.
- we recommend measuring reaction immediately but reaction is stable for at least 2 hours.
1) Prepare reaction mix with Assay Buffer 23, OxiRed Probe, Converter Mix B and Developer Mix N, and add to sample and standard wells.
Omit Converter Mix B from reaction mix added to sample background wells. In absence of Converter Mix B, the assay detects only endogenous glycerol phosphate but not ATP.
2) Mix and incubate at room temperature for 30 min protected from light.
3) Measure output on a microplate reader at OD 570 nm for colorimetric assay, or at Ex/Em = 535/587 nm for fluorometric assay.
Publications (517)
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