AST/GOT enables alpha-ketoglutaric acid and aspartic acid to displace amino and keto groups to form glutamic acid and oxaloacetic acid. Oxaloacetic acid can decarboxylate itself to form Pyroracemic acid during the reaction. Pyroracemic acid reacted with 2,4-dinitrophenylhydrazine(DNPH) to form 2,4, dinitrophenylhydrazone showing reddish brown in alkaline solution. Measure the OD values and calculate the enzyme activity.
AST can catalyze the substrate to produce pyruvate. Pyruvate is oxidized to produce hydrogen peroxide, which causes the fluorescent probe to produce fluoresce. The activity of AST can be calculated by measuring the increase of fluorescence value at the excitation wavelength of 535 nm and the emission wavelength of 587 nm.
The detection principle of this kit is that the enzyme catalyzes aspartic acid and substrate to produce glutamic acid, and glutamic acid is further catalyzed by the enzyme to produce a product, which will be reduced to orange yellow product under the action of electronic coupler, and the maximum absorption peak is detected at 450 nm.
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