PhosphoWorks™ Colorimetric ATP Assay Kit

PhosphoWorks™ Colorimetric ATP Assay Kit is a colorimetric assay kit for quantifying ATP levels in biological samples.

Detection Principle: ATP-driven enzyme reactions generate H₂O₂ for colorimetric detection.
Sensitivity: Detects approximately 3 µM ATP in a 100 µL reaction.
Specificity: Shows minimal interference from ADP and AMP.
Workflow: Incubates 10–30 minutes at room temperature.
Instrument Compatibility: Absorbance detection at 570 nm using clear-bottom plates.

ATP dose response measured with PhosphoWorks&trade; Colorimetric ATP Assay Kit in a 96-well clear bottom plate using a SpectraMax microplate reader (Molecular Devices).

Protocol

SDS

COA

Catalog	Size	Price	Quantity
21617	100 Tests	Price

Overview

Adenosine triphosphate (ATP) plays a fundamental role in cellular energetics, metabolic regulation and cellular signaling. It is referred as the "molecular unit of currency" of intracellular energy transfer to drive many processes and chemical synthesis in living cells. ATP also serves as a signaling molecule for cell communication and plays an important role in DNA and RNA synthesis. AAT Bioquest offers a variety of bioluminescence assay kits to determine nanomolar (nM) range of ATP with recombinant firefly luciferase (Cat# 21610 & 21609). These kits require luminescence plate readers, are frequently used for cell viability or cytotoxicity assays. PhosphoWorks™ Colorimetric ATP Assay Kit is based on a serial ATP-induced enzyme coupled reactions to produce hydrogen peroxide, which is spectrophotometrically quantified with our Amplite® Red Substrate at OD 570 nm. The assay can detect ~3 µM of ATP in a 100 µL reaction volume with minimal interference from ADP and AMP. It provides a robust, simple and convenient assay for measuring ATP levels in biological samples. The PhosphoWorks™ Colorimetric ATP Assay is complementary to our luciferase-based ATP assay kits.

Citations

View all 50 citations: Citation Explorer

Photothermal-Responsive Conjugated Polymer Nanoparticles Triggered Precise Augmentation of Microalgal Photosynthesis

Authors:

Zhang, Miaomiao and Gao, Chenying and Wang, Pengcheng and Li, Rui and Huang, Yiming and Lv, Fengting and Li, Cheng and Bai, Haotian and Zhang, Deqing and Wang, Shu

Journal:

ACS nano (2025)

PERIOD 2 Regulates Low Dose Radioprotection via PER2/pGSK3$\beta$/$\beta$-Catenin/Per2 Loop

Authors:

Alexandrou, Aris T and Duan, Yixin and Xu, Shanxiu and Tepper, Clifford and Fan, Ming and Tang, Jason and Berg, Jonathan and Basheer, Wassim and Valicenti, Tyler and Wilson, Paul F and others,

Journal:

Iscience (2022): 105546

ATP-based cell viability assay is superior to trypan blue exclusion and XTT assay in measuring cytotoxicity of anticancer drugs Taxol and Imatinib, and proteasome inhibitor MG-132 on human hepatoma cell line HepG2

Authors:

Nowak, E., Kammerer, S., Kupper, J. H.

Journal:

Clin Hemorheol Microcirc (2018): 327-336

High-content image analysis (HCIA) assay has the highest correlation with direct counting cell suspension compared to the ATP, WST-8 and Alamar blue assays for measurement of cytotoxicity

Authors:

Tahara, H., Matsuda, S., Yamamoto, Y., Yoshizawa, H., Fujita, M., Katsuoka, Y., Kasahara, T.

Journal:

J Pharmacol Toxicol Methods (2017): 92-99

High throughput cell-based assay for identification of glycolate oxidase inhibitors as a potential treatment for Primary Hyperoxaluria Type 1

Authors:

Wang, Mengqiao and Xu, Miao and Long, Yan and Fargue, Sonia and Southall, Noel and Hu, Xin and McKew, John C and Danpure, Christopher J and Zheng, Wei

Journal:

Scientific Reports (2016)

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	570 nm
Recommended plate	Clear bottom

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Page updated on October 28, 2026

Component A: Amplite® Red Substrate (light sensitive)	1 vial
Component B: Enzyme Mix	1 bottle (Lyophilized powder)
Component C: Assay Buffer	1 bottle (5 mL)
Component D: ATP Standard	2.8 mg/vial
Component E: DMSO	1 vial (100 µL)