Application of Biological Buffer HEPES 7365-45-9 in Lactate Dehydrogenase Detection Kit

As a key indicator in clinical diagnosis and biological research, the stability and accuracy of lactate dehydrogenase (LDH) detection kits directly affect the reliability of experimental results. Among the various components of the reagent kit, the biological buffer 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES buffer) has become a core element in maintaining pH stability in the detection environment due to its unique physicochemical properties.

Chemical properties and advantages of HEPES

HEPES is a zwitterionic biological buffering agent, whose molecular structure includes a piperazine ring, hydroxyethyl, and sulfonic acid groups, endowing it with excellent buffering ability. Its pKa value is 7.55, close to the physiological pH value, exhibiting efficient buffering performance in the pH range of 6.8-8.2, especially suitable for biological reactions under neutral to weakly alkaline conditions. Compared with traditional buffering agents such as Tris and phosphate, HEPES has the following significant advantages:

hepes powder

1. Low adsorption: Its surface inertness can reduce non-specific adsorption of biomolecules such as proteins and enzymes, avoiding experimental errors;

2. Temperature stability: not easily decomposed at conventional laboratory temperatures, suitable for long-term experiments and sample preservation;

3. Biocompatibility: It is non-toxic to cells, does not affect enzyme activity or cell metabolism, and meets the safety requirements of biological testing.

In LDH detection, the reagent kit needs to simulate the intracellular environment in vitro. The above characteristics of HEPES can ensure the pH stability of the reaction system, avoiding enzyme activity distortion or sample degradation caused by pH fluctuations.

The mechanism of HEPES in LDH detection kit

The core principle of LDH detection is to reduce NAD+to NADH through enzymatic reaction, which reacts with chromogenic agents (such as INT) to generate measurable formaldehyde products. During this process, slight changes in pH may directly affect enzyme activity, substrate binding efficiency, and color reaction rate. HEPES ensures the accuracy of detection through the following mechanisms:

1. PH stability: In the reaction system of the reagent kit, HEPES buffer can neutralize the hydrogen ions or hydroxide ions generated during the reaction process, maintain a constant pH, and ensure that the enzymatic reaction proceeds under optimal conditions;

2. Anti interference: Its low ion strength characteristic can reduce the interference of impurities (such as salts and proteins) in the sample on the reaction, and improve the detection sensitivity and specificity;

3. Extended shelf life: During the storage stage of the reagent kit, HEPES can prevent enzyme or color reagent inactivation caused by pH drift and extend the validity period of the reagent.

Taking a lactate dehydrogenase cytotoxicity detection kit as an example, its reaction system includes HEPES buffer, lactate substrate, NAD+, and color reagent. During the detection process, HEPES buffer (pH 7.5) can stably maintain the enzymatic reaction environment, ensuring a linear relationship between the generation rate of NADH and the color intensity of formaldehyde products, thereby accurately quantifying LDH activity through colorimetric methods.

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Hubei Xindesheng Material Technology Co., Ltd. specializes in producing high-quality biological buffering agents such as Hepes. Since its establishment, Desheng has been dedicated to the research and production of in vitro diagnostic reagents and has rich production experience. Our professional technical team not only ensures product quality, but also provides customized solutions and timely resolution of after-sales issues. If you are interested in our products, please click on the official website to learn more details or contact me!