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Cat. No.: F6PW8-100 (for 100T)

Description

Fructose-6-phosphate Assay Kit with WST-8, also known as the 6-Phosphate Fructose Assay Kit (WST-8 method) or F6P Assay Kit with WST-8, is a kit based on the WST-8 chromogenic reaction. This kit utilizes a colorimetric method to quickly and sensitively detect fructose-6-phosphate content in biological fluids such as tissue or cell samples, serum and plasma, as well as tissue or cell culture supernatants.

Fructose-6-phosphate (F6P), also known as 6-Phosphate Fructose, has the molecular formula C6H13O9P and a molecular weight of 260.13. It is a phosphorylated product of fructose. F6P is a significant metabolic intermediate in cellular metabolism. During glycolysis, glucose-6-phosphate (G6P) is isomerized into F6P by phosphoglucose isomerase (PGI). F6P plays a role in initiating glycogen synthesis and is involved in the hexosamine pathway and the hexose monophosphate shunt. Additionally, F6P also participates in the non-oxidative branch of the pentose phosphate pathway (PPP), producing pentose phosphates used for nucleotide synthesis and erythrose 4-phosphate, a precursor for various aromatic metabolites and histidine biosynthesis in aromatic amino acid autotrophs.

Principle

Fructose-6-phosphate (F6P) is isomerized into glucose-6-phosphate (G6P) by phosphoglucose isomerase (PGI). The generated G6P is oxidized by glucose-6-phosphate dehydrogenase (G6PDH) into 6-phosphogluconate (6-PG), reducing NADP+ to NADPH in the process. NADPH, in the presence of an electron mediator, reduces WST-8 to orange-yellow formazan, which has a maximum absorbance peak around 450 nm. The amount of formazan produced in the reaction system is proportional to the fructose-6-phosphate content in the sample.

Components

• Buffer A for Metabolic Assay
• F6P Assay Buffer
• Substrate
• Enzyme Solution A
• Enzyme Solution B
• WST-8
• F6P Standard (100mM)

Feature

• This assay kit features high sensitivity, a wide linear range, and requires a small sample volume. With a sample volume of 20µl, this kit can detect fructose-6-phosphate concentrations as low as 5μM. It demonstrates a good linear relationship within the concentration range of 5-2000μM (0.1-40nmol). The kit provides a fructose-6-phosphate standard solution, which allows for the creation of a standard curve to calculate the fructose-6-phosphate content in samples.
• The detection lysis buffer provided in this kit has a certain universality. The cell or tissue samples lysed using Buffer A for Metabolic Assay from this kit can also be used for sample detection in other metabolic assay kits produced by Beyotime that use Buffer A for Metabolic Assay. This provides strong versatility. Furthermore, it can also be used for protein concentration detection, SDS-PAGE, or Western detection of some easily soluble proteins.
• This assay kit is flexible, fast, and widely applicable. It can be used for the detection of biological fluids such as serum, plasma, and urine from mice, rats, and humans, as well as cell culture supernatants and tissue or cell samples. The entire process can be completed in approximately 0.5-1 hour. This kit is suitable not only for the detection of small sample quantities but also for high-throughput screening and automated systems.

Storage

Store at -20ºC for one year. WST-8 must be stored protected from light.

Precautions

• Buffer A for Metabolic Assay, F6P Assay Buffer, and Substrate need to be completely thawed and equilibrated to room temperature before use; otherwise, it may affect the detection results. Other solutions should be used on ice.
• Substrate may precipitate upon thawing from -20ºC. The precipitate will re-dissolve once equilibrated to room temperature, which will not affect the detection results.
• Serum and other samples stored at 4ºC should not be stored for more than two weeks, as this may affect the accuracy of detection results. Generally, serum samples should be stored at -20ºC, with -80ºC being preferable.
• This product is intended for scientific research by professionals only. It is not to be used for clinical diagnosis or treatment, food, or medicine, nor should it be stored in ordinary residences.
• For your safety and health, please wear a lab coat and disposable gloves when operating.

Only for research and not intended for treatment of humans or animals

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