Exploring the Mysteries of High purity Biobuffer HEPES CAS 7365-45-9

Lactate dehydrogenase (LDH), as one of the key enzymes in the intracellular glycolysis pathway, undergoes changes in its activity under various pathological conditions. Accurately measuring LDH activity is not only of great significance for basic scientific research, but also an important tool for clinical diagnosis. Choosing the appropriate buffer is one of the key factors in ensuring the accuracy of the test results during this process.

Hydroxyethyl piperazine ethanesulfonic acid (HEPES buffer), as a widely used biological buffer, is gradually becoming the core component of lactate dehydrogenase detection kits. This article will analyze in detail the key role of HEPES in lactate dehydrogenase detection from multiple perspectives, including its importance and application in LDH detection.

HEPES powder

The importance of HEPES in lactate dehydrogenase detection

The redox reaction between lactate and pyruvate catalyzed by LDH requires a specific pH range to exhibit optimal activity. When the pH deviates from the optimal value, the activity of the enzyme may decrease or even be completely lost, resulting in biased detection results. Therefore, adding buffering agents to the detection system of lactate dehydrogenase is a key step to ensure detection accuracy.

HEPES plays a crucial role in lactate dehydrogenase detection kits due to its excellent buffering performance. In the process of LDH catalyzed reduction of pyruvic acid to L-lactic acid, this is a reversible reaction, and when the concentration of lactic acid is high, it promotes the reverse reaction, that is, the synthesis of pyruvic acid from lactic acid. Under suitable pH conditions maintained in HEPES buffer, L-lactic acid reacts with NAD+to produce pyruvic acid and NADH. NADH exhibits characteristic absorbance at a wavelength of 340nm, which means that LDH activity can be quantified by monitoring changes in absorbance. Therefore, HEPES not only maintains the pH stability of the reaction system, but also provides a guarantee for accurate determination of lactate dehydrogenase content.

Application of HEPES in lactate dehydrogenase detection kit

The reagent kit used for lactate dehydrogenase detection usually consists of independent reagents I and II.

In reagent I, HEPES acts as a biological buffer and forms a reaction system with metal ion complexes, LDH substrates, surfactants, LDH enzyme activators, preservatives, and water. HEPES is mainly responsible for maintaining the pH stability of the reaction system and providing a suitable reaction environment for the substrate of lactate dehydrogenase. At the same time, it interacts with metal ion complexes to ensure that metal ions (such as zinc ions in lactate dehydrogenase) are in a stable chemical state, maintaining the enzyme's active conformation. In addition, surfactants, enzyme activators, and other components can better promote reactions in the stable pH environment created by HEPES.

In reagent II, HEPES also exists and works together with nicotinamide adenine dinucleotide oxidation state (NAD ⁺), surfactants, preservatives, and water. NAD ⁺, as a coenzyme in lactate dehydrogenase catalyzed reactions, is highly sensitive to pH in the redox reactions it participates in. HEPES can ensure that NAD ⁺ synergizes with lactate dehydrogenase under suitable pH conditions to complete the substrate redox process. In addition, HEPES also works together with preservatives to maintain the stability of reagent II and extend the shelf life of the reagent kit.

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Hubei Xindesheng Material Technology Co., Ltd. specializes in producing high-quality hepaes and other biological buffering agents. Our professional technical team ensures product quality and can also provide customized solutions to timely after-sales problems. If you are interested in our products, please click on the official website to learn more details or contact me!