[92a] Microsomal lipid peroxidation

doi:10.1016/0076-6879(67)10099-2

Methods in Enzymology

Volume 10, 1967, Pages 574-580

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This chapter discusses the microsomal lipid peroxidation that may be demonstrated by measuring (1) O₂ consumption, (2) NADPH disappearance, and (3) malonaldehyde (MA) formation. Results of an experiment involving the measurement of all three parameters are illustrated. In the experimental procedure, rat liver microsomes are prepared s by sedimenting the 10,000 g supernatant of a 0.25M sucrose homogenate of rat liver at 105,000 g for 60 minutes. O₂ consumption is measured with a Clark O₂ electrode. NADPH disappearance is monitored fluorometrically at 450 mμ with an excitation wavelength of 365mμ. An Eppendorf photometer with fluorometer attachment is a suitable instrument for this purpose. MA formed is measured colorimetrically with the thiobarbituric acid (TBA) reaction. O₂ consumption may alternatively be measured manometrically in the Warburg apparatus. Measurement of NADPH disappearance spectrophotometrically at 340 mμ is complicated by the turbidity of the microsomal suspension which, in addition, may change during the incubation. The maximal rate of NADPH-linked lipid peroxidation at 30° is approximately 160 millimicromoles of O₂ consumed per minute per milligram of microsomal protein. The NADPH disappearance accompanying the microsomal lipid peroxidation ranges between one-third and one-fifth mole of NADPH per mole of O₂ consumed. The maximal extent of lipid peroxidation is approximately one micromole of O₂ consumed per milligram of microsomal protein.

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[92a] Microsomal lipid peroxidation

Publisher Summary

Biochem. Biophys. Res. Commun.

Nature

Biochem. Biophys. Res. Commun.

Biochem. Pharmacol.

Anal. Biochem.

J. Biol. Chem.

J. Biol. Chem.

Food. Res.

Arch. Biochem. Biophys.

J. Biol. Chem.

J. Biol. Chem.

J. Cell Biol.