A standard curve was prepared using 100 units/mL of SOD enzyme solution (Sigma-Aldrich, MO, USA) as a standard compound. SOD levels were determined by adding the supernatant into the reaction cocktail (pH 7.8) containing 0.108 mM xanthine solutions (Sigma-Aldrich, MO, USA), 216 mM potassium phosphate buffer (pH 7.8, Ajax Finechem, Auckland, New Zealand), 1.1 mM cytochrome C (Sigma-Aldrich, MO, USA), and 10.7 mM ethylenediaminetatraacetic acid (Sigma-Aldrich, MO, USA). After that, the reaction mixture was reacted with 0.1 units/mL of xanthine oxidase enzyme solution (Sigma-Aldrich, MO, USA) before analyzing the absorbance at 540 nm at 0 and 5 min in triplicate.
Xanthine oxidase enzyme solution
Manufactured by Merck Group
Sourced in United States
Xanthine oxidase enzyme solution is a laboratory reagent produced by Merck Group. It contains the enzyme xanthine oxidase, which catalyzes the oxidation of xanthine and hypoxanthine to uric acid. The solution is intended for use in biochemical and enzymatic applications where the activity of xanthine oxidase is required.
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Lab products found in correlation
2 protocols using xanthine oxidase enzyme solution
1
SOD Enzyme Quantification Assay
2
Evaluating Anti-Gout Potential via Xanthine Oxidase Inhibition
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In vitro xanthine oxidase inhibition assay was used to evaluate anti-gout potential of the plant. To prepare plant extract solution, extract (2 mg) was dissolved in 1 ml 5% DMSO in water solution. In this mixture, 2.9 ml phosphate buffer (1.15 M, pH=7.5) and 0.1 ml of xanthine oxidase enzyme solution (purchased from Sigma) were added. After 15 minutes of mixture pre incubation at 25 0 C temperature, the reaction was started by adding a substrate solution (2 ml).Then after 30 minutes, the reaction was stopped by adding 1 ml of IN HCl. Absorbance was measured at 290nm. Likewise, blank was prepared but enzyme was being added to assay mixture after addition of the HCl. Xanthine oxidase activity was calculated by following formula and expressed as the percentage inhibition of xanthine oxidase. % Inhibition of xanthine oxidase = 1-(A extract/A no extract) x 100 Where, A is the absorbance (Guerrero et al., 2011) .
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