Lactate dehydrogenase activity (WST-8 method) microcalorimetry

Lactate dehydrogenase activity (WST-8 method) microcalorimetryPrinciple of MeasurementIt's through testingLDHDuring the process of catalyzing the reduction of pyruvic acid to lactic acidNADHTo indirectly determine the amount of generationLDHThe activity.

The specific process is as follows:

Reaction principle:inLDHUnder its influence,NAD+Generated by restorationNADH.NADHandWST-8The interaction generates color, which is then used for colorimetric measurement (λ)max=450 nm）.

Reaction conditions:LDHTranslate into EnglishNADRestore toNADH，NADHandWST-8Interaction produces color. This reaction takes place on ice in the dark, with each component (such as...)Assay BufferTheLactateTheNADTheWST-8TheEnhancer）It needs to be removed from the refrigerator and equilibrated to room temperature before the experiment, and the entire process should be carried out in the dark.

Absorption measurement:in450 nmMeasure absorbance at wavelength, absorbance andLDHThe activity is proportional and can be determined by colorimetric methodLDHThe activity.

Experimental steps and required equipment:

Experimental steps:Prepare working fluid（1×Assay BufferTheNADTheWST-8TheEnhancerandLactateMix evenly), add the working fluid96Add the sample to be tested into an orifice plate or micro glass colorimetric dish,37Measure the absorbance after incubating at a constant temperature of ℃ for a certain period of time.

Required equipment:ELISA reader or visible spectrophotometer (capable of measuring)450 nmThe absorbance of the sample, constant temperature box, ice maker, low-temperature centrifuge96Orifice plate or micro glass colorimetric dish, adjustable pipette and tip, deionized water, homogenizer (if it is a tissue sample).

Experimental precautions‌：

Reagent storage:Each component (such asAssay BufferTheLactateTheNADTheWST-8TheEnhancer）It needs to be stored on ice in the dark to avoid repeated freezing and thawing.

Precautions for operation:The entire experimental process needs to be conducted on ice in the dark to avoid light affecting the results.

Lactate dehydrogenase activity (WST-8 method) microcalorimetryThe products currently being operated by the company:

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