As an index of lipid peroxidation, we used the formation of thiobarbituric acid-reactive species (TBARS) by measuring the amount of MDA, formed from the breakdown of polyunsaturated fatty acids during an acid heating reaction, as described previously (Ohkawa et al. 1979) (link). Briefly, the supernatants were mixed with 0.67% (w/v) thiobarbituric acid (Merck) and heated in a boiling water bath for 20 min. After cooling, the absorbance of each sample was determined at 535 nm using a Beckman Coulter u.v./Visible (DU800). TBARS content was expressed in micromoles of MDA formed per milligram of protein.
U v visible du800
Manufactured by Beckman Coulter
The Beckman Coulter U.v./Visible (DU800) is a spectrophotometer that measures the absorbance of light in the ultraviolet and visible spectrum. It is designed to analyze the concentration and purity of various samples in a laboratory setting.
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Lab products found in correlation
2 protocols using u v visible du800
1
Lipid Peroxidation Assessment via TBARS
2
Enzymatic Glutathione Quantification
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GSH content was determined by an enzymatic assay as described previously (Tietze 1969) (link). Briefly, the supernatants were incubated with 0.24 mmol/l NADPH, 0.75 mmol/l 5,5 0 -dithiobis-2-nitrobenzoic acid in PBS 0.05 M, pH 7.4, and 2.5 IU GSH reductase, and the development of the chromophoric product was recorded every 30 s for 6 min at an absorbance of 412 nm using a Beckman Coulter u.v./Visible (DU800). Total GSH content was expressed in micromoles GSH per milligram of protein.
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