Amplite® Colorimetric Fructose-6-Phosphate Activity Assay Kit

Amplite® Colorimetric Fructose-6-Phosphate Activity Assay Kit is a colorimetric assay kit for quantifying fructose-6-phosphate (F6P) through an enzyme-coupled NADH readout in a microplate format.

Colorimetric detection: Measures an NADH-dependent colored product proportional to F6P concentration.
Coupled enzyme chemistry: Links F6P conversion through PFK, aldolase, and GAPDH reactions to NADH production.
Rapid workflow: Incubate for 10–30 minutes at room temperature before reading.
Microplate compatible: Read absorbance at 450 nm using a clear-bottom plate.
Applications: Suitable for F6P quantitation in glycolysis and cellular metabolism research.

The dose response of F6P was measured using the Amplite® Colorimetric Fructose-6-Phosphate Assay Kit. The assay was conducted on a 96-well clear bottom microplate, with incubation for 30 minutes, and readings were taken at 450 nm using a ClarioStar microplate reader (BMG).

Protocol

SDS

COA

Catalog	Size	Price	Quantity
11330	100 Tests	Price

Overview

The Amplite® Colorimetric Fructose-6-Phosphate Assay Kit is a highly sensitive and simple method of quantifying F6P present in a variety of samples. This kit uses a coupled enzyme assay where F6P, in the presence of ATP, is converted to fructose-1,6-diphosphate and ADP by the PFK enzyme. The aldolase enzyme then converts fructose-1,6-diphosphate into dihydroxyacetone phosphate and glyceraldehyde-3-phosphate. Subsequently, glyceraldehyde-3-phosphate dehydrogenase, in the presence of NAD, converts glyceraldehyde-3-phosphate into glyceraldehyde-1,3-bisphosphate and NADH. The NADH produced then reduces a colorless substrate to a colored product (A450 nm), which is proportional to the fructose-6-phosphate present. One unit (U) is the amount of enzyme that catalyzes the reaction of 1 µmol of substrate per minute. Fructose-6-phosphate (F6P) is an important intermediate generated during glycolysis. It is produced by the isomerization of glucose-6-phosphate by phosphoglucose isomerase. It serves as a precursor to fructose 1,6-bisphosphate. Beyond its role in energy production, F6P can also be shunted to the non-oxidative pentose phosphate pathway as a means of generating pentose phosphates for nucleotide production. F6P levels surge in rapidly proliferating cells like cancer cells, making it a potential metabolic biomarker in diseases.

References

View all 48 references: Citation Explorer

Overexpression of Fatty Acid Synthase Upregulates Glutamine-Fructose-6-Phosphate Transaminase 1 and O-Linked N-Acetylglucosamine Transferase to Increase O-GlcNAc Protein Glycosylation and Promote Colorectal Cancer Growth.

Authors:

Drury, James and Geisen, Mariah E and Tessmann, Josiane Weber and Rychahou, Piotr G and Kelson, Courtney O and He, Daheng and Wang, Chi and Evers, B Mark and Zaytseva, Yekaterina Y

Journal:

International journal of molecular sciences (2024)

Silencing of Glutamine: Fructose-6-Phosphate Aminotransferase Impairs Growth and Development in Sogatella furcifera (Hemiptera: Delphacidae).

Authors:

Wang, Zhao and Long, Guiyun and Zhu, Huan and Jin, Daochao and Yang, Hong and Zhou, Cao

Journal:

Biomolecules (2023)

Dry-milled flour rice 'Seolgaeng' harbors a mutated fructose-6-phosphate 2-kinase/fructose-2,6-bisphosphatase2.

Authors:

Kang, Ju-Won and Lee, Sang-Kyu and Shim, Su-Hyeon and Shin, Dongjin and Cho, Jun-Hyeon and Lee, Ji-Yoon and Ko, Jong-Min and Ji, Hyeonso and Park, Hyang-Mi and Ahn, Eok-Keun and Lee, Jong-Hee and Jeon, Jong-Seong

Journal:

Frontiers in plant science (2023): 1231914

Cancer-associated fibroblast expression of glutamine fructose-6-phosphate aminotransferase 2 (GFPT2) is a prognostic marker in gastric cancer.

Authors:

Yang, Shuo and Li, Guoli and Yin, Xin and Wang, Yufei and Jiang, Xinju and Bian, Xiulan and Fang, Tianyi and Yin, Shengjie and Zhang, Lei and Xue, Yingwei

Journal:

The journal of pathology. Clinical research (2023): 391-408

Fructose-6-phosphate-2-kinase/fructose-2,6-bisphosphatase regulates energy metabolism and synthesis of storage products in developing rice endosperm.

Authors:

Chen, Xiaoli and Ji, Yi and Zhao, Weiying and Niu, Huanying and Yang, Xue and Jiang, Xiaokang and Zhang, Yipeng and Lei, Jie and Yang, Hang and Chen, Rongbo and Gu, Chuanwei and Xu, Hongyi and Dong, Hui and Duan, Erchao and Teng, Xuan and Wang, Yunlong and Zhang, Yuanyan and Zhang, Wenwei and Wang, Yihua and Wan, Jianmin

Journal:

Plant science : an international journal of experimental plant biology (2023): 111503

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 26, 2026

Component A: F6P Probe	1 Vial (1 mL)
Component B: F6P Assay Buffer	1 Bottle
Component C: F6P Enzyme Mix	1 Bottle
Component D: F6P Developer	1 Bottle
Component E: ATP	1 Bottle (4 mL)
Component F: Fructose-6-Phosphate (F6P) Standard	1 Vial