Amplite® Colorimetric Phosphofructokinase (PFK) Activity Assay Kit

Amplite® Colorimetric Phosphofructokinase (PFK) Activity Assay Kit is a colorimetric assay kit for quantifying phosphofructokinase activity in biological samples.

Detection Principle: Uses a coupled enzymatic reaction to convert PFK activity into a measurable colorimetric signal.
Sensitivity: Enables quantitative detection of changes in PFK activity through signal intensity proportional to enzyme activity.
Reaction Chemistry: Uses fructose-6-phosphate and ATP as substrates in the PFK-catalyzed reaction.
Instrument Compatibility: Designed for absorbance measurement at 450 nm using a microplate reader.
Applications: Supports studies of glycolysis and metabolic enzyme activity in biological samples.

NADH dose response was measured using the Amplite® Colorimetric Phosphofructokinase (PFK) Assay Kit. The measurements were taken on a 96-well clear bottom microplate after a 30-minute incubation period using a ClarioStar microplate reader (BMG) at 450 nm.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
11329	100 Tests	Price

Overview

The Amplite® Colorimetric Phosphofructokinase (PFK) Activity Assay Kit provides a simple and direct procedure for precise measurement of PFK enzyme activity in various biological samples. This kit is based on a coupled enzyme assay, in which fructose-6-phosphate in the presence of ATP is converted to fructose1,6-diphosphate and ADP by PFK enzyme. This ADP is then converted to AMP and NADH by an enzyme mix. The resulting NADH reduces a colorless substrate to a colored product which absorbs at 450 nm and is proportional to the PFK activity present. One unit (U) is the amount of enzyme that catalyzes the reaction of 1 µmol of substrate per minute. Phosphofructokinase (PFK) is an enzyme of glycolysis pathway catalyzing the conversion of fructose-6-phosphate to fructose-1,6-diphosphate. This is a crucial step of glycolysis as this reaction determines the overall rate of glycolysis. PFK enzyme activity is regulated by multiple cofactors and post-translational modifications. Deficiencies in PFK activity are linked to a rare genetic disorder known as glycogen storage disease type VII (GSD-VII), also known as Tarui disease. Additionally, deficiencies in PFK activity may also contribute to exercise-induced myopathy and rhabdomyolysis in certain individuals.

References

View all 50 references: Citation Explorer

Disruption of phosphofructokinase activity and aerobic glycolysis in human bronchial epithelial cells by atmospheric ultrafine particulate matter.

Authors:

Park, Su Hwan and Kim, Gyuri and Yang, Gi-Eun and Yun, Hye Jin and Shin, Tae Hwan and Kim, Sun Tae and Lee, Kyuhong and Kim, Hyuk Soon and Kim, Seok-Ho and Leem, Sun-Hee and Cho, Wan-Seob and Lee, Jong-Ho

Journal:

Journal of hazardous materials (2024): 132966

Biological and Genetic Determinants of Glycolysis: Phosphofructokinase Isoforms Boost Energy Status of Stored Red Blood Cells and Transfusion Outcomes.

Authors:

Nemkov, Travis and Stephenson, Daniel and Earley, Eric J and Keele, Gregory R and Hay, Ariel and Key, Alicia and Haiman, Zachary and Erickson, Christopher and Dzieciatkowska, Monika and Reisz, Julie A and Moore, Amy and Stone, Mars and Deng, Xutao and Kleinman, Steven and Spitalnik, Steven L and Hod, Eldad A and Hudson, Krystalyn E and Hansen, Kirk C and Palsson, Bernhard O and Churchill, Gary A and Roubinian, Nareg and Norris, Philip J and Busch, Michael P and Zimring, James C and Page, Grier P and D'Alessandro, Angelo

Journal:

bioRxiv : the preprint server for biology (2024)

Retraction notice to "Phosphofructokinase-M inhibits cell growth via modulating the FOXO3 pathway in renal cell carcinoma cells" [Biochemical and Biophysical Research Communications 530(2020) 67-74].

Authors:

Chen, Yeda and Yu, Qingfeng and Duan, Xiaolu and Wu, Wenqi and Zeng, Guohua

Journal:

Biochemical and biophysical research communications (2024): 149619

Phosphofructokinase family genes in grass carp: Molecular identification and tissue-specific expression in response to glucose, insulin and glucagon.

Authors:

Zhang, Yingxin and Qin, Chaobin and Wang, Junli and Yang, Liping and Yan, Xiao and Zhi, Shaoyang and Nie, Guoxing

Journal:

Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology (2024): 110898

Structural basis for allosteric regulation of human phosphofructokinase-1.

Authors:

Lynch, Eric M and Hansen, Heather and Salay, Lauren and Cooper, Madison and Timr, Stepan and Kollman, Justin M and Webb, Bradley A

Journal:

bioRxiv : the preprint server for biology (2024)

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	450 nm
Recommended plate	Clear bottom

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Page updated on October 16, 2026

Component A: PFK Probe	1 Vial (1 mL)
Component B: PFK Assay Buffer	1 Bottle (4 mL)
Component C: PFK Enzyme Mix	1 Bottle
Component D: PFK Developer	1 Vial
Component E: ATP	1 Vial
Component F: NADH Standard	1 Vial
Component G: PFK Positive Control	1 Vial