Isolation of intracellular membranes by means of sodium carbonate treatment: Application to Endoplasmic Reticulum

Yukio Fujiki, L. Hubbard, Stanley Fowler, Paul B. Lazarow

Research output: Contribution to journalArticlepeer-review

Abstract

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 Na2CO3 followed 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. (1974, J. Cell Biol. 61: 213-231). 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 (1982, J. Cell Biol. 93: 103-110) the procedure is applied to peroxisomes and mitochondria.

Original languageEnglish (US)
Pages (from-to)97-102
Number of pages6
JournalJournal of Cell Biology
Volume93
Issue number1
DOIs
StatePublished - Apr 1 1982

ASJC Scopus subject areas

  • Cell Biology

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