Quantification of Intracellular Metabolites

Analyzing the various intracellular metabolisms i.e., the glycolysis system, the tricarboxylic acid (TCA) cycle, and the electron transport chain, is important when trying to understand cellular states. Metabolites and energy sources [e.g., glucose, lactate, and NAD (P)/NAD (P) H] are the indicators used for the analysis of intracellular metabolisms.

Amerigo Scientific offers metabolic assay kits that include all necessary components for sample preparation and measurement.

Easy Procedure for First Time User

For a first-time user, the kit includes the reagents and components necessary for measuring samples.

Determination index	Detection	Operation
Glucose Glutamine Glutamate Lactate NAD/NADH NADP/NADPH	Colorimetric	Simply transfer the culture supernatant to a plate and mix it with the chromogenic reagent Easy pretreatment with the included protein removal tube* and cell lysis buffer
ATP	Luminescent	Kit includes ATP standard - very easy to use
α-ketoglutaric acid	Fluorescent	Less variable results than existing assays

Products Range

•  Glucose and Lactate Assay Kits

Glucose, the most important biological energy source, is a known marker for energy metabolism. It is used not only as a marker for diabetes and obesity but also for monitoring changes in intracellular metabolism. Due to its reliability in detecting metabolic changes, its application has been seen in many different fields of study, including cancer research.

Lactate quantification has been widely used as a marker for monitoring changes in intracellular metabolic pathways in many different areas of cancer research. In recent years, there have been an increasing number of publications that have used lactate as an indicator for stem cell differentiation or as an indicator in diabetic research.

•  NAD+/NADH and NADP+/NADPH Assay Kits

Nicotinamide adenine dinucleotide (NAD) is an important cofactor involved in redox reactions in the major cellular metabolic pathways such as glycolysis, the electron transfer system, and the TCA cycle. NAD exists in cells in an oxidized form, NAD+, and a reduced form, NADH. Maintaining appropriate NAD+ and NADH levels is essential for cellular function. In addition, a recent study suggests that a decrease in NAD+ levels is associated with cellular senescence.

Nicotinamide adenine dinucleotide phosphate (NADP) is an important cofactor involved in the pentose phosphate pathway reactions (another of the cellular metabolic pathways). NADP exists in cells in an oxidized form as NADP+ and in a reduced form as NADPH. NADP+ and NADPH are not only involved in the biosynthesis of fatty acids and cholesterol but also in the generation of reduced glutathione (GSH), an important antioxidant. Recent studies suggest that carbohydrate restriction can lead to shortened life expectancy, as consequence of an increase in the NADP+/NADPH ratio, which leads to decreased GSH and thus increased DNA damage by reactive oxygen species.

•  Glutamine and Glutamate Assays

Glutamine, a source of α-ketoglutarate, a TCA-cycle intermediate, is an important substance used for energy production, as well as the synthesis of nucleic acids and other amino acids. Glutaminolysis, the process by which glutamine as a substrate is converted to α-ketoglutarate, is upregulated in cancer cells.

Glutaminase Electron mediator Glutamate is an important neurotransmitter that contributes to the biosynthesis of protein and glutathione. Excess glutamate is considered to be a cause of neurodegenerative disorders, such as Alzheimer's disease.

•  α-Ketoglutarate and ATP Assays

α-Ketoglutarate (α-KG) is an important intermediate of the TCA cycle. It is used as an indicator of both activity in the glutaminolysis pathway, and the uptake of glucose metabolites into the TCA cycle. α-KG plays an essential role in the production of glutamate and γ-aminobutyric acid (GABA) and contributes to the scavenging of reactive oxygen species.

Adenosine triphosphate (ATP) is an energy source for living cells, with 95% of cellular ATP synthesized in mitochondria. Because of this, ATP levels can generally be considered an indicator of mitochondrial activity. Decreased ATP levels are associated with mitochondrial diseases, as well as neurodegenerative diseases, cancer, and aging.