Biophysical Journal 64: 1063-1068 (1993)
© 1993 the Biophysical Society

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Polyethylene glycol as a hydration agent in oriented membrane bilayer samples

Department of Physics, University of British Columbia, Vancouver, British Columbia, V6T 1Z1, Canada

ABSTRACT

Techniques such as NMR, ESR, fluorescence depolarization, and neutron scattering are commonly used to investigate the physical properties of membranes. Oriented membrane bilayer systems (single crystals) are often employed in these investigations. It is important to know and be able to control the level of hydration in these samples. In particular, one must have confidence that a sample is in fact "fully hydrated" and remains so during the course of the experiment. Full hydration is difficult to obtain by hydrating oriented samples using water-saturated vapor. An alternative method for hydrating oriented samples is to surround the oriented sample by a polymer solution. Higher hydration levels are achieved using this method. Three nuclear magnetic resonance studies using headgroup deuterated 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were done to compare the hydration level of oriented headgroup samples surrounded by a polymer/water solution and fully hydrated multibilayer dispersions. Transition temperatures, quadrupolar splittings (at 50°C) and spin-lattice relaxation times (at 50°C) were measured. The simple tests of the transition temperature and quadrupolar splitting to determine full hydration, as my results show, are not sufficient. In this paper I demonstrate that more fully hydrated samples can easily be achieved by surrounding the oriented sample with a 5 wt% polyethylene glycol/water solution than by hydrating in water saturated vapor.