Determination of Significance:
ehydroascorbate reductase (DHAR) is an important antioxidant enzyme in plants and a key enzyme that promotes ascorbic acid regeneration in the ascorbate-glutathione oxidation cycle. In the circulation DHAR maintain the normal metabolic level of ascorbic acid in plants through ascorbic acid, and plays an important role in protecting cellular components from oxidative damage.
Measurement Principle:
DHAR catalyzes the reduction of dehydroascorbic acid (DHA) by reducing glutathione (GSH) to produce AsA. GSH can react with 5,5'-dithio-bis- (2-nitrobenzoic acid) (DTNB) to produce 2- Nitro-5-mercaptobenzoic acid (TNB) and glutathione disulfide (GSSG). TNB has maximum light absorption at a wavelength of 412 nm. DHAR activity is calculated by measuring the reduction rate of GSH.
Self Provided:
Low temperature centrifuge, spectrophotometer, water bath, mortar/homogenizer, 1 mL glass cuvette, adjustable pipette, ice and distilled water.
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