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Purification of Plasma Membrane from Acanthamoeba castellanii 1
Author(s) -
CLARKE BARBARA J.,
HOHMAN THOMAS C.,
BOWERS BLAIR
Publication year - 1988
Publication title -
the journal of protozoology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 0022-3921
DOI - 10.1111/j.1550-7408.1988.tb04118.x
Subject(s) - percoll , membrane , chromatography , chemistry , plasma , biochemistry , centrifugation , physics , quantum mechanics
A simple method for isolation of plasma membrane from Acanthamoeba using self‐generating gradients of Percoll is described. To obtain a membrane marker, intact amoebae were radioiodinated and the distribution of the radiolabel was followed through the plasma membrane isolation procedure. The purity of isolated plasma membrane was assessed by enrichment of radiolabel, by electron microscopy, and by enzymatic assays for contaminating membranes. As judged from enrichment of radiolabel, a 37‐fold purification of plasma membrane was obtained. We estimate that 80% of the total protein was from plasma membrane and 10% from membrane‐associated actin.