CheKine™ Micro α-Ketoglutarate Dehydrogenase (α-KGDH) Assay Kit

α-Ketoglutarate Dehydrogenase (α-KGDH) is widely present in mitochondria of the animal, plant, microorganism and in-vitro cultured cells. It is one of the key regulation enzyme of Kreb's Cycle, and can catalyze α-Ketoglutarate oxidation and decarboxylation to produce succinyl coenzyme A.

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• simple, convenient, rapid and colorimetric α-KGDH activity detection method. • compatible to various biological samples, especially animals or plant tissues or cell cultures (adherent or suspension) and fungi .• Provides detailed sample preparation and results calculation methods.

CheKine™ Micro α-Ketoglutarate Dehydrogenase (α-KGDH) Assay Kit provides a simple, sensitive, rapid and colorimetric α-KGDH activity detection method, which is compatible to various biological samples, especially animals or plant tissues or cell cultures (adherent or suspension) and fungi. The principle of detection is that α-KGDH can catalyze α-Ketoglutarate, NAD + and coenzyme A to produce succinyl-coenzyme A, carbon dioxide and NADH. NADH has a characteristic absorption peak at 340 nm. α-KGDH activity could be calculated according to NADH-generating rates.

• ΔA of Test Tube should be between 0.01-0.3, the sample should be diluted further if ΔA of Test Tube > 0.3. •Do not mix the components between different batch numbers and manufacturers; otherwise, the results may be abnormal. • Avoid bubbles while mixing or redissolving components. •Change pipette tips frequently to avoid cross contamination between components. • Ensure that all components and equipment are at the proper temperature before starting the experiment. • All samples and reagents should be on ice to avoid denaturation and deactivation . • Don't recommend to test too many samples in one time , because enzyme activity is calculated by the variation of the absorbance value per unit time . •Recommended to calculate the enzyme activity by protein concentration. If you need to calculate by the sample weight , the total enzyme activity is equal to the enzyme activity of the supernatant and pellet which is cytoplasmic extraction. •Please to extract α-KGDH from fresh samples in order to ensure the enzyme activity . • Please reduce the tissues weight if the OD values is too high . Or reduce the volume of Extraction Buffer if the OD values is too low .