[Rapid simultaneous isolation of microsomes and plasma membranes from neuroblastoma C 1300 N 18 cells].
The method is suggested to isolate simultaneously microsomes and plasma membranes of neuroblastoma S 1300 N 18 cells by means of differential centrifugation in the step density gradient of Percoll/Ficoll with a high degree of purification determined from the activity of marker enzymes (acetyl cholinesterase Na+,K+-ATPase, alkali phosphatase, glucose-6-phosphatase, succinate-dehydrogenase, acid phosphatase) as well as from the content of DNA and RNA and with a sufficiently high protein yield. The purified fractions of microsomes and plasma membranes are established to contain no phosphatidyl glycerol and cardiolipin--safety markers of mitochondrial membrane purification. A degree of separation of microsomes, plasma membranes and proteins dissolved in cytosol may be estimated by the activity of the cholesterol-synthesizing system of enzymes with the use of sterol-transferring protein.
Volkov
G L
GL
D000818:Animals; D002308:Cardiolipins / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure*; D002498:Centrifugation; D004273:DNA, Neoplasm / Q000032:analysis; D004798:Enzymes / Q000378:metabolism; D051379:Mice; D008861:Microsomes / Q000032:analysis / Q000648:ultrastructure*; D009363:Neoplasm Proteins / Q000032:analysis; D009447:Neuroblastoma / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure*; D010715:Phosphatidylglycerols / Q000032:analysis; D012334:RNA, Neoplasm / Q000032:analysis; D014407:Tumor Cells, Cultured / Q000648:ultrastructure
D002308:Cardiolipins; D004273:DNA, Neoplasm; D004798:Enzymes; D009363:Neoplasm Proteins; D010715:Phosphatidylglycerols; D012334:RNA, Neoplasm
Bystroe odnovremennoe vydelenie mikrosom i plazmaticheskikh membran kletok neĭroblastomy C 1300 N 18.