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. 1982 Sep 1;205(3):511–519. doi: 10.1042/bj2050511

Isolation of plasma membrane from protoplasts of Lolium multiflorum (ryegrass) endosperm cells.

A Schibeci, G B Fincher, B A Stone, A B Wardrop
PMCID: PMC1158515  PMID: 7150229

Abstract

Plasma membranes have been isolated from protoplasts of suspension-cultured ryegrass (Lolium multiflorum) endosperm cells. The protoplast membrane is coated before cell disruption with murine myeloma protein J539, a galactose-binding immunoglobulin A. The plasma membrane is labelled with 125I by using chemically or enzymically catalysed iodination techniques, or, more conveniently, by using 125I-labelled myeloma protein J539, which enables the membrane to be simultaneously coated and labelled. Protoplast lysis is effected by gentle mechanical means after swelling in hypo-osmotic medium. The plasma-membrane fraction is recovered at low centrifugal forces by fractionation of cell lysates on a discontinuous sucrose/sorbitol gradient. The plasma-membrane fraction is enriched 96-fold on a protein basis with respect to the specific radioactivity of 125I-labeled myeloma protein J539 in the homogenate. Electron microscopy showed long membrane profiles often associated with one another.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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