Plasma membranes from fibroblastic cells in culture isolation,morphological and enzymatic identification |
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Authors: | Lionel Lelievre |
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Institution: | Unité d''Immunodifférenciation, Laboratoire de Microscopie Electronique, Institut de Biologie Moléculaire, 2 Place Jussieu, 75-Paris 5ème, France |
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Abstract: | Plasma membranes were isolated from murine plasmocytoma cells in culture, by a procedure involving lysis in hypoosmotic medium leaving the nuclei intact, and separation of surface membranes from the lysate constituents on a discontinuous sucrose gradient.The purity of the fractions was assessed by electron microscopy and by assaying enzymes for cross-contaminants. Phosphohydrolases, including the (Na+ + K+)-stimulated Mg2+-ATPase (EC 3.6.1.3) and 5′-nucleotidase (EC 3.1.3.5), were concentrated in the plasma membrane-rich fractions. These fractions were essentially free from NADH: cytochrome c reductase, lysosomes and mitochondrial membrane enzymes. |
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