By Arthur Weissbach and Herbert Weissbach (Eds.)

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Leonard, this series, Vol. 32, p. 392. 2 (4ml/g tissue). 8 SUCROSE GRADIENT centrifuge 95 kg 2h f 20% 30% 34% ΕΣΧΧΣ PM 45% PHASE PARTITION preweighed| PEG Θ dextran mixture centrifuge at 1kg for 5 min supernatant (discard) pellet, resuspend in grinding medium y resuspend upper phase and lower phase (with the interphase) in 10 volumes of grinding medium PM MU other u · Ί interphase centrifuge at 150 kg for 6 0 min membranesl FIG. 1. Flow chart depicting the steps involved in the purification of plasma membranes using either a sucrose gradient or an aqueous two-phase separation.

Chanson, E. McNaughton, and L. Taiz, Plant PhysioL 76, 498 (1984). 28 P. M. Ray, in "Plant Organelles" (E. ), pp. 135-146. Ellis Horwood, Chichester, 1979. 24 [2] ISOLATION OF THE PLASMA MEMBRANE 47 Mitochondria, cytochrome c oxidase119 Endoplasmic reticulum, NADH cytochrome c reductase, antimycin A insensitive1929 130 Golgi vesicles, IDPase glucan synthetase I 130 Tonoplast, NOf-inhibited ATPase3132 (still uncertain) None of the plasma membrane markers, when used alone, is totally satisfactory.

Harris, R. J. Henry, A. B. Blakeney, and B. A. Stone, Carbohydr. Res. 127, 59 (1984). A. J. Mort, S. -S. Kuo, Anal. Biochem. 133, 380 (1983). [1] CELL WALLS 35 tridge. ) The per-O-methylated polysaccharide is then eluted from the Sep-Pak C-18 cartridge with a 2-mlflushof 100% acetonitrile and, after that, a 4-ml flush of 100% ethanol. The 100% acetonitrile and 100% ethanol flushes, containing the per-O-methylated polysaccharide, are collected in a test tube and the solvent is evaporated to dryness under a stream offilteredair at room temperature.

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