As a commonly used sub-Kelvin refrigeration technology, helium sorption coolers play an important role in space and ground applications. The adsorption characteristics of the porous material inside the sorption cooler at low temperature have a crucial influence on its performance. At present, the analysis and calculation of sorption coolers are mainly based on helium 4 (4He) as the working gas, and there is a lack of systematic research on the low-temperature adsorption characteristics of helium-3 (3He) and its coupling effect characteristics of temperature, pressure, and mass distribution in different components. In this paper, a molecular model of activated carbon that is similar to the actual structure was constructed, and the adsorption isobars and isosteric heat of 3He and 4He at 0.8–5 K were comparatively studied based on the grand canonical Monte Carlo (GCMC) method. Besides, the influence of adsorption characteristics of 3He and 4He on the condensation efficiency, the mass distribution after condensation equilibrium, and the self-cooling loss of the sorption cooler were analyzed. The results show that for the 3He sorption cooler, the main factor affecting the condensation efficiency is the adsorbed helium in the sorption pump, while for the 4He sorption cooler, it is the adsorbed helium and the gas in the dead volume. For both 3He and 4He sorption coolers, the condensation efficiency increases as the sorption pump temperature increases or the heat sink temperature decreases, while the self-cooling loss decreases as the heat sink temperature decreases or the operating temperature increases.