The plasma membrane from suspended-cultured Populus euphratica(PE) cells had been isolated and purified by two-phase partition composed of Dextran T-500 and PEG 3350. Effect of different concentrations of the polymer (5.5%，5.7%，5.9%，6.1%，6.3%，6.5%) and KCl (0，5，10，15 mmol/L) on the purification were examined. The results indicated that two-phase system composed of 5.9% polymer without salt resulted in plasma membrane of PE with higher H^＋-ATPase activity. The activity of the membrane H^＋-ATPase has been increased 8-fold (from 4.23 to 32.63 μmol/mg·h) with better orientation and most of the membrane H^＋-ATPases are right side-out. These results may provide better basis and availability for further studies of the purified H^＋-ATPase from PE plasma membrane.