PhosphoWorks™ Fluorimetric ATP Assay Kit

PhosphoWorks™ Fluorimetric ATP Assay Kit is a fluorimetric assay kit for quantifying ATP in biological samples using an enzyme-coupled hydrogen peroxide readout.

Detection Principle: Converts ATP through coupled enzymatic reactions to generate H₂O₂.
Sensitivity: Detects approximately 0.4 µM ATP in a 100 µL reaction.
Specificity: Shows minimal interference from ADP and AMP.
Chemistry: Uses Amplite® Red for fluorescent H₂O₂ detection.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm using solid black plates.

ATP dose response was measured with PhosphoWorks&trade; Fluorimetric ATP Assay Kit in a 96-well solid black plate using a Germini microplate reader (Molecular Devices).

Protocol

SDS

COA

Catalog	Size	Price	Quantity
21620	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™ Fluorimetric 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. The assay can detect ~0.4 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™ Fluorimetric ATP Assay is complementary to our luciferase-based ATP assay kits.

Citations

View all 49 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)

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)

NT1014, a novel biguanide, inhibits ovarian cancer growth in vitro and in vivo

Authors:

Zhang, Lu and Han, Jianjun and Jackson, Am and a L , undefined and Clark, Leslie N and Kilgore, Joshua and Guo, Hui and Livingston, Nick and Batchelor, Kenneth and Yin, Yajie and Gilliam, Timothy P and others, undefined

Journal:

Journal of Hematology & Oncology (2016): 91

References

View all 15 references: Citation Explorer

Fluorescent detection of apoptotic cells by using zinc coordination complexes with a selective affinity for membrane surfaces enriched with phosphatidylserine

Authors:

Hanshaw RG, Lakshmi C, Lambert TN, Johnson JR, Smith BD.

Journal:

Chembiochem (2005): 2214

Detection of apoptotic cells using a synthetic fluorescent sensor for membrane surfaces that contain phosphatidylserine

Authors:

Koulov AV, Stucker KA, Lakshmi C, Robinson JP, Smith BD.

Journal:

Cell Death Differ (2003): 1357

Interactions of a vesicular stomatitis virus G protein fragment with phosphatidylserine: NMR and fluorescence studies

Authors:

Hall MP, Burson KK, Huestis WH.

Journal:

Biochim Biophys Acta (1998): 101

Influence of annexin V on the structure and dynamics of phosphatidylcholine/phosphatidylserine bilayers: a fluorescence and NMR study

Authors:

Saurel O, Cezanne L, Milon A, Tocanne JF, Demange P.

Journal:

Biochemistry (1998): 1403

A Chinese hamster ovary cell mutant defective in the non-endocytic uptake of fluorescent analogs of phosphatidylserine: isolation using a cytosol acidification protocol

Authors:

Hanada K, Pagano RE.

Journal:

J Cell Biol (1995): 793

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	540 mn
Emission	590 nm
Cutoff	570 nm
Recommended plate	Solid black

Telephone
Fax
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Technical support	Contact us
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Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Page updated on October 22, 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)