Isolation of Intracellular Membranes by Means of Sodium Carbonate Treatment: Application to Endoplasmic Reticulum

A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content protein...

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Bibliographic Details
Published in:The Journal of cell biology 1982-04, Vol.93 (1), p.97-102
Main Authors: Fujiki, Yukio, Hubbard, Ann L., Fowler, Stanley, Lazarow, Paul B.
Format: Article
Language:English
Subjects:
Animals
Carbonates
Cell Fractionation - methods
Cell membranes
Centrifugation, Density Gradient - methods
Cytochromes
Female
Intracellular Membranes - ultrastructure
Liver
Liver - ultrastructure
Membrane proteins
Microscopy, Electron
Microsomes
Microsomes, Liver - ultrastructure
P branes
Rats
Rats, Inbred Strains
Ribosomes
RNA
Sarcoplasmic Reticulum - ultrastructure
Sodium
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Description
Summary:A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content proteins and peripheral membrane proteins are released in soluble form. Here we document the method by applying it to various subfractions of a rat liver microsomal fraction, prepared by continuous density gradient centrifugation according to Beaufay et al. The results confirm and extend those of previous investigators on the distribution of enzymes and proteins among the membranes of the smooth and rough endoplasmic reticulum. In the accompanying paper the procedure is applied to peroxisomes and mitochondria.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.93.1.97