PhosphoWorks™ Fluorimetric Phosphate Assay Kit *Red Fluorescence*

PhosphoWorks™ Fluorimetric Phosphate Assay Kit

Red Fluorescence

PhosphoWorks™ Fluorimetric Phosphate Assay Kit *Red Fluorescence* is a fluorimetric assay kit for detecting inorganic phosphate (Pi) and measuring the activity of Pi-generating enzymes in a microplate format.

Detection Principle: Uses a red fluorescent phosphate sensor responsive to Pi.
Sensitivity: Quantifies phosphate down to 0.1 µM.
Applications: Supports phosphatase, ATPase, and GTPase activity measurements.
Workflow: Compatible with 96- or 384-well assays and can be easily adapted to automation with no separation steps.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm using solid black plates.

Phosphate dose response was measured with PhosphoWorks™ Fluorimetric Phosphate Assay Kit on a solid black 96-well plate using a Novostar microplate reader (BMG Labtech).

Protocol

Catalog	Size	Price	Quantity
21660	125 Tests	Price
21661	1250 Tests	Price

Overview

Cells utilize a wide variety of phosphate (Pi) and polyphosphate esters as enzyme substrates, second messengers, membrane structural components and vital energy reservoirs. Phosphate is involved in many biological processes. For example, phosphatases, ATPases and several other enzymes catalyze biochemical reactions in which inorganic phosphate is released from a phosphoester substrate. Detection of many phosphoester-metabolizing enzymes is difficult because suitable substrates are not available. It usually has been necessary to determine inorganic phosphate release using tedious colorimetric assays or radioisotope-based methods. This PhosphoWorks™ Fluorimetric Phosphate Assay Kit has been developed for measuring the activity of any Pi-generating enzyme using our red fluorescent phosphate sensor. The kit provides sensitive detection of Pi, an alternative to hazardous radioactive methods and other less sensitive colorimetric assays. The measurement of Pi is based on the change in the absorbance and fluorescence of our new phosphate sensor. Our kit provides all the essential reagents including phosphate sensor, phosphate standards and assay buffer. The assay is shown to quantitate phosphate in solution at concentrations at least down to 0.1 µM. It can be used to measure the kinetics of phosphate release from phosphatases (such as GTPases and ATPases) by coupling the two enzymatic reactions. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation with no separation steps required.

Citations

View all 8 citations: Citation Explorer

Whole-genome sequencing reveals an East Asian-specific rare variant of INPP5J associated with Alzheimer’s disease

Authors:

Kimura, Tetsuaki and Yamakawa, Akiko and Mitsumori, Risa and Niida, Shumpei and Ozaki, Kouichi and Shigemizu, Daichi

Journal:

Translational Psychiatry (2026)

Pseudomonas aeruginosa lasR-deficient mutant contributes to bacterial virulence through enhancing the PhoB-mediated pathway in response to host environment

Authors:

Pan, Xiaolei and Yin, Liwen and Zhou, Dandan and Jin, Yongxin and Cheng, Zhihui and Ha, Un-Hwan and Jin, Shouguang and Wu, Weihui

Journal:

mBio (2025): e01788--25

Pseudomonas aeruginosa Phosphate Transporter PitA (PA4292) Controls Susceptibility to Aminoglycoside Antibiotics by Regulating the Proton Motive Force

Authors:

Zhao, Xinrui and Jin, Yongxin and Bai, Fang and Cheng, Zhihui and Wu, Weihui and Pan, Xiaolei

Journal:

Antimicrobial Agents and Chemotherapy (2022): e00992--22

Bladder epithelial cell phosphate transporter inhibition protects mice against uropathogenic Escherichia coli infection

Authors:

Pang, Yu and Cheng, Zhihui and Zhang, Si and Li, Shujie and Li, Xueping and Li, Xiaodan and Zhang, Xiao and Li, Xiaoxiao and Feng, Yingxing and Cui, Heting and others,

Journal:

Cell Reports (2022): 110698

Sacrificial crystal templating of hyaluronic acid-based hydrogels

Authors:

Thomas, Richelle C and Chung, Paul E and Modi, Shan P and Hardy, John G and Schmidt, Christine E

Journal:

European Polymer Journal (2017): 487--496

References

View all 8 references: Citation Explorer

Relationship between activating and editing functions of the adenylation domain of apo-tyrocidin synthetase 1 (apo-TY1)

Authors:

Bucevic-Popovic V, Pavela-Vrancic M, Dieckmann R, Von Dohren H.

Journal:

Biochimie (2006): 265

Purine nucleoside phosphorylase activity in rat cerebrospinal fluid

Authors:

Silva RG, Santos DS, Basso LA, Oses JP, Wofchuk S, Portela LV, Souza DO.

Journal:

Neurochem Res (2004): 1831

Characterization of the interactions between the small GTPase RhoA and its guanine nucleotide exchange factors

Authors:

Tan YC, Wu H, Wang WN, Zheng Y, Wang ZX.

Journal:

Anal Biochem (2002): 156

A continuous spectrophotometric assay for aspartate transcarbamylase and ATPases

Authors:

Rieger CE, Lee J, Turnbull JL.

Journal:

Anal Biochem (1997): 86

Continuous monitoring of Pi release following nucleotide hydrolysis in actin or tubulin assembly using 2-amino-6-mercapto-7-methylpurine ribonucleoside and purine-nucleoside phosphorylase as an enzyme-linked assay

Authors:

Melki R, Fievez S, Carlier MF.

Journal:

Biochemistry (1996): 12038

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

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

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Email	sales@aatbio.com
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Page updated on October 28, 2026

Component A: Assay Buffer	1 bottle (5 mL)
Component B: Phosphate Sensor	1 vial (lyophilized powder)
Component C: 1 mM KH2PO4	1 vial (1 mL)

Component A: Assay Buffer	10 Bottles
Component B: Phosphate Sensor	10 vials (lyophilized powder)
Component C: 1 mM KH2PO4 Standard	1 vial (1 mL)