Plasma membranes of human neutrophils: a one-step isolation procedure by cell disruption in paraffin oil.
Plasma membranes of high purity and good yield have been prepared from human polymorphonuclear neutrophils by a one-step procedure involving disruption of cells suspended in paraffin oil and forced by pressure through an annular slit. This results in a band floating above the oil which is composed of large sheets of plasma membranes. Enrichment values for the plasma membrane marker alkaline phosphatase and 125I-labeled protein after surface labeling performed at the whole cell level were 23-fold and 22-fold, respectively. Contamination of the plasma membrane by other organelles was negligible and approximately 2 mg of membrane protein was obtained from 10(9) neutrophils. The procedure is very fast and the use of paraffin oil avoids lengthy high-speed centrifugation. The technique also allows isolation of granules devoid of plasma membrane and can probably be applied to other cell types.
D000328:Adult; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000201:enzymology; D003593:Cytoplasm / Q000032:analysis; D006801:Humans; D008565:Membrane Proteins / Q000097:blood; D009504:Neutrophils / Q000032:analysis / Q000648:ultrastructure*; D009821:Oils; D010232:Paraffin; D013347:Subcellular Fractions / Q000032:analysis
D008565:Membrane Proteins; D009821:Oils; D010232:Paraffin; C015418:paraffin oils