Preparation of guinea pig hepatic microsomes: a comparison of three techniques.
Hepatic microsomal preparations by Ca2+-aggregation and gel filtration on Sepharose 2B were compared with the product obtained by differential centrifugation, using liver from adult male guinea pigs. The conventionally-prepared microsomes were translucent and rose-red in color. Those prepared by Ca2+-aggregation were a cream or light pink color whereas the gel-filtered microsomes were opalescent and white. The specific activities of microsomal p-nitroanisole O-demethylase, aniline hydroxylase and nonspecific carboxylase were highest in the Ca2+-aggregated microsomes, intermediate in the gel-filtered preparation and lowest in the conventional microsomes. Electron microscopy revealed vesicular preparations of similar appearance though the Ca2+-aggregated microsomes were larger and contained more aggregates of free ribosomes.
D000815:Aniline Hydroxylase / Q000378:metabolism; D000818:Animals; D002122:Calcium Chloride / Q000494:pharmacology; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002850:Chromatography, Gel; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008297:Male; D008722:Methods; D008854:Microscopy, Electron; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D009575:Nitroanisole O-Demethylase / Q000378:metabolism; D014461:Ultracentrifugation
D009575:Nitroanisole O-Demethylase; D000815:Aniline Hydroxylase; D002265:Carboxylic Ester Hydrolases; D002122:Calcium Chloride