Equilibrium and kinetic aspects of Triton X-100 adsorption onto hydrophobic Bio-Beads SM2 were investigated in detail using the batch procedure originally described by Holloway, P.W. (1973) Anal. Biochem. 53, 304-308. The results demonstrated the importance of the initial detergent concentration, the amount of beads, the commercial source of the detergent, the temperature and the presence of phospholipids in determining the rates of Triton X-100 adsorption onto Bio-Beads. One of the main findings was that Bio-Beads allowed the almost complete removal of Triton X-100, whatever the initial experimental conditions. It was shown that monomeric as well as micellar detergent could be adsorbed and that a key factor in determining the rate of detergent removal was the availability of the free bead surface. Rates of detergent removal were found to be linearly related to the amount of beads even for bead concentrations above those sufficient to remove all the detergent initially present. Adsorptive capacity of phospholipids onto Bio-Beads SM2 was also analyzed and found to be much smaller (2 mg lipid per g of wet beads) than that of Triton X-100 (185 mg TX 100 per g of wet beads). A more general aspect of this work was that the use of Bio-Beads SM2 provided a convenient way for varying and controlling the time course of Triton X-100 removal. The method was further extended to the formation of liposomes from phospholipid-Triton X-100 micelles and the size of the liposomes was found to be critically dependent upon the rate of detergent removal. A general procedure was described to prepare homogeneous populations of vesicles. Freeze-fracture electron microscopy and permeability studies indicated that the liposomes thus obtained were unilamellar, relatively large and impermeable. Noteworthy, this new procedure was shown to be well suited for the reconstitution of different membrane transport proteins such as bacteriorhodopsin, Ca2(+)-ATPase and H(+)-ATPase.