Name: ATP Assay Kit (Colorimetric/Fluorometric)
SKU: ab83355
Rating: 4 (5 reviews)

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Key features and details

Assay type: Quantitative
Detection method: Colorimetric/Fluorometric
Platform: Microplate reader
Assay time: 1 hr
Sample type: Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
Sensitivity: 1 µM

Product name

ATP Assay Kit (Colorimetric/Fluorometric)
See all ATP kits
Detection method

Colorimetric/Fluorometric
Sample type

Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate
Assay type

Quantitative
Sensitivity

< 1 µM
Assay time

1h 00m
Product overview

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

Chinese protocol available. See protocols section below.

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.
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.

Related assays

Review the cell health assay guide to learn about kits to perform a cell viability assay, cytotoxicity assay and cell proliferation assay.

Review the metabolism assay guide 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}
Platform

Microplate reader

Storage instructions

Store at -20°C. Please refer to protocols.

Components	100 tests	2000 tests
Assay Buffer XXIII	1 x 25ml	20 x 25ml
ATP Standard	1 vial	20 vials
Converting Enzyme I	1 vial	20 vials
Developer IV	1 vial	20 vials
OxiRed™ Probe	1 x 200µl	20 x 200µl

Research areas
Alternative names
- Adenosine 5' triphosphate

Corresponding kit
- Deproteinizing Sample Preparation Kit - TCA (ab204708)

ATP levels in Pancreatic IsletImage courtesy of Mrs. Fotini Mouth

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.
ATP assay used to study mitochondrial disfunction after C. rodentium infection in miceImage courtesy of Maiti A K et al. PLoS One. 2018; 13(9): e0204567. doi: 10.1371/journal.pone.0204567

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.
ATP assay performed with ab83355Sanokawa-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/

The chart shows a comparison of ATP levels of HepG2 treated with 0, 10 and 100 µM for 48 hours, DR^{H2O2 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).
ATP assay performed with ab83355Image 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/

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 10⁵cells per well and incubated for two days under standard culture conditions. ATP levels are measured using ATP assay kit (ab83355).
ATP assay performed with ab83355

Example of fluorometric ATP assay standard curve.
ATP levels measured in mouse colon tissueImage 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/

Maiti AK et al. (2015) used ATP assay kit ab113852 to measure mitochondrial ATP generation in murine distal colon after C. rodentium infection.
ATP assay performed with ab83355

Example of colorimetric ATP assay standard curve.
ATP assay performed with 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).

Protocol Booklet

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab83355? Please let us know so that we can cite the reference in this datasheet.

ab83355 has been referenced in 373 publications.

Vu Hong A et al. Dlk1-Dio3 cluster miRNAs regulate mitochondrial functions in the dystrophic muscle in Duchenne muscular dystrophy. Life Sci Alliance 6:N/A (2023). PubMed: 36265896
Rybkowska P et al. The Metabolic Changes between Monolayer (2D) and Three-Dimensional (3D) Culture Conditions in Human Mesenchymal Stem/Stromal Cells Derived from Adipose Tissue. Cells 12:N/A (2023). PubMed: 36611971
Woo JH et al. Polypropylene nanoplastic exposure leads to lung inflammation through p38-mediated NF-κB pathway due to mitochondrial damage. Part Fibre Toxicol 20:2 (2023). PubMed: 36624477
Piazzesi A et al. CEST-2.2 overexpression alters lipid metabolism and extends longevity of mitochondrial mutants. EMBO Rep 23:e52606 (2022). PubMed: 35297148
He L et al. An Effective Sodium-Dependent Glucose Transporter 2 Inhibition, Canagliflozin, Prevents Development of Hypertensive Heart Failure in Dahl Salt-Sensitive Rats. Front Pharmacol 13:856386 (2022). PubMed: 35370704

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1-5 of 5 Abreviews

We tested the kit ab83355 to quantify the total ATP in 80 hours post fertilization zebrafish larvae. ATP was quantified by fluorometric method (Ex/Em = 535/587 nm) using a TECAN Infinite 200 PRO.
Ten l0 hours post fertilization (hpf) larvae were prepared following the protocol given by Abcam. Two technical replicates were run in parallel with a background control (mix without ATP converter).

According to our results, the background noise generate by glycerol phosphate does not allow the quantification of ATP in the larvae. Additionally, the kit seems to be not sensitive enough to detected ATP in 80 hpf zebrafish larvae. In fact, all measured values are below the last point of the calibration curve.

We tried different experimental set ups:
Fig. 1: Deproteinization using PCA (45 minutes) as described by the Abcam protocol. Each bar represents a biological replicate. 50 μl of sample was tested.
Fig. 2: Different deproteinization times (5 and 1 minute) and no deproteinization step. 25 μl of sample was tested.
Fig.3: Different dilutions (10 larvae in 100 μl buffer and 50 μl buffer) and clean-up without using acids (chloroform clean-up). 40 and 30 μl of sample were tested, respectively. Experiments were performed in duplicates.

Despite we tried different dilutions, deproteinization and clean-up steps, the general low concentration of ATP and the strong background noise does not allow the quantification of ATP in the larvae. Consequently, we do not recommend to use this kit for zebrafish larvae.
However, we would happy to try a more sensitive methods such as the Luminescent ATP Detection Assay Kit (ab113849).