2-DG is up-taken by the cells, converted to 2-DG-6P, which is catalyzed by glucose dehydrogenase to produce 6PDG. Meanwhile, NADP+ is converted to NADPH. The generated NADPH converts the probe into fluorescent substances under the action of myocardial yellow transferase. The glucose uptake can be calculated by measuring the fluorescence intensity at the excitation wavelength of 530 nm and the emission wavelength of 590 nm.
GDH catalyzes the dehydrogenation of glutamate, meanwhile, NAD+ is reduced to NADH, Which under the action of hydrogen transmitter, transfer electrons to WST-8 to produce the yellow product. The activity of GDH can be calculated by measuring the change of absorbance value at 450 nm.
GOGAT catalysis the reaction that transfer the amino from glutamine to a-KG to form two molecules of glutamic acid using NADH as the electron donor. The decreasing rate of NADH that can be measured at 340 nm can reflect the activity of GOGAT.
Glutamic acid can react with NAD+ under the catalysis of glutamate dehydrogenase to produce α-ketoglutaric acid, NADH and NH4+. NADH has the maximum absorption at 340 nm. And glutamic acid content can be calculated by measuring the change of NADH.
Glutamic acid can reduce NAD+ to NADH. NADH, under the action of hydrogen transmitter, reduces WST-8 to form yellow product, which has a characteristic absorption peak at 450 nm. Glutamic acid content can be calculated by measuring the OD value at 450 nm.
Glutamine is decomposed to produce glutamic acid under the action of glutaminase. Glutamic acid is further transformed by glutamic acid dehydrogenase. Meanwhile, NAD+ is reduced to NADH, Which under the action of hydrogen transmitter, transfer electrons to WST-8 to produce the yellow product. The activity of glutaminase can be calculated by measuring the change of absorbance value at 450 nm.
Gln is hydrolyzed to produce glutamic acid under the action of glutaminase. Glutamic acid is further catalyzed by glutamic acid dehydrogenase. Meanwhile, NAD+ is reduced to NADH, Which under the action of hydrogen transmitter, transfer electrons to WST-8 to produce the yellow product. The content of Gln can be calculated by measuring the change of absorbance value at 450 nm.
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