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• W2000935328 abstract "Epithelial cells from isolated rat small intestine were harvested by vibration of the everted intestine in 0.14 m NaCl containing 5 mm EDTA. These cells, which were largely free of mucus contamination, were homogenized in hypotonic (74 mm) sucrose using a Potter-Elvehjem homogeniser. After successively sedimenting a “brush border plus nuclei” and a “mitochondrial” fraction, microsomes were prepared from the postmitochondrial supernatant by ultracentrifugation or by precipitation at pH 5.0. The isolation and fractionation procedure was validated by the distribution of marker enzymes and by light microscopy and found to be largely uncontaminated by other subcellular components or by haemoglobin. The usefulness of this preparation was demonstrated by determining drug-metabolising enzyme activity and by substrate- and metabolite-binding spectra to cytochrome P-450. A comparison of precipitated “acid” and “normal” intestinal microsomes indicated similar apparent Km and Vmax values for a number of drug-metabolising enzymes. The content of components of the microsomal electron transport system were also similar in both preparations while the “acid” microsomes contained approximately 50% more protein." @default.
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• W2000935328 date "1979-01-01" @default.
• W2000935328 modified "2023-10-04" @default.
• W2000935328 title "An improved method for preparing rat small intestine microsomal fractions for studying drug metabolism" @default.
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• W2000935328 doi "https://doi.org/10.1016/s0003-2697(79)80118-9" @default.
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