Coenzyme I NAD(H) Content Assay Kit (MTT, Micro Method)

Cat. No.: C1527139

GRADE & PURITY BioReagent

Overview

Coenzyme I NAD(H) is widely present in animals, plants, microorganisms, and cultured cells. NAD⁺ serves as the primary hydrogen acceptor in glycolysis (EMP) and the tricarboxylic acid (TCA) cycle. The NADH generated from its reduction transfers electrons to oxygen via the electron transport chain (ETC), producing ATP while generating a large amount of reactive oxygen species (ROS), and NADH is regenerated to NAD⁺. The vast majority of oxidative reactions involved in the catabolism of carbohydrates, lipids, and proteins proceed through this system. Therefore, the NAD(H) content and the NADH/NAD⁺ ratio reflect the intensity of glycolysis and the TCA cycle: an increased ratio suggests higher cellular oxygen consumption and an oxidative stress state, which in turn exerts feedback inhibition on glycolysis and the TCA cycle. Additionally, degradation products of NAD⁺ play important regulatory roles in cellular signaling, metabolic regulation, and gene expression. This kit employs acidic and alkaline extraction buffers to separately extract NAD⁺ and NADH from samples. NADH reduces oxidized thiazolyl blue tetrazolium bromide (MTT) to formazan via the electron-carrying action of phenazine methosulfate (PMS), and the absorbance is measured at 570 nm. NAD⁺ is first reduced to NADH by alcohol dehydrogenase, followed by quantification through the aforementioned MTT reduction system, thereby enabling the separate quantification of NAD⁺ and NADH.

Storage and Shipping

Storage

Store at 2-8°C

Shipped In

Wet ice