AbstractIn Part I of this paper, a thermoelectrogravitational electrophoresis column without reservoirs was proposed for preparative electrophoretic separations. The Furry, Jones, and Onsager procedure in thermal diffusion was applied to develop a transport equation, and its solution for the steady state batch case was employed to demonstrate some typical effects of temperature difference, electric field strength, and membrane spacing on the predicted steady state batch separation in the column.In this part, a theory is first developed from mathematical analysis of a continuous‐flow thermoelectrogravitational column without reservoirs by modification of the transport equation to take into account the bulk flow through the column.An experimental center‐fed thermoelectrogravitational electrophoresis column and the related equipment used to obtain experimental data to test the theory are described. Further, experimental separation factors as a function of flow rates ranging from 0 to 10 g/min for the bovine albumin system at twopH values (8.6 and 6.0) were obtained using two membrane spacings (0.1354 and 0.3018 cm respectively) at four different electric field strengths (0.0423 to 0.423 volt/cm) and for three different temperature differences of 0°C, 8.5°C, and 16°C. Experimental data indicated that meaningful separations could be obtained using thermoelectrogravitational columns but that the temperature difference has an adverse effect on electrophoretic separation.Analysis of the experimental flow data showed that theory and experiment are not in quantitative agreement. However, there was general, qualitative agreement between theory and experiment for the dependence of separation on field strength, temperature difference, membrane spacing and mobility of the com