Chapter 14 The Plasma Membrane of KB Cells; Isolation and Properties

Abstract

Publisher Summary This chapter describes the isolation of the plasma membrane of KB cells by two different methods, namely, the latex bead ingestion and the ZnCl 2 methods. In principle, the latex bead method can be employed with any cell type provided the proper conditions for bead ingestion are optimized (size and multiplicity of beads, incubation medium, time and temperature of incubation, and absence of fusion between phagosomes and lysosomes). The method has been used for the isolation of phagosomes from Acanthamoeba, L cells, and polymorphonuclear leukocytes, and macrophages. Optimal conditions are developed for the ingestion of latex beads by mouse neuroblastoma cells (clone Neuro-2A) in monolayer (differentiated state) and suspension cultures (undifferentiated form). It is observed that only lymph node macrophages ingest latex beads (1.01 μm in diameter) after intradermal-subcutaneous injection of a 10% aqueous suspension of beads. Thus the isolation of plasma membranes by the latex bead procedure described can be accomplished with cells grown in culture as well as with certain cells from tissues after in vivo injection of the beads.