Gas adsorption on the surface of nanoporous rocks is an important process that occurs in many applied scenarios such as shale gas production or CO2 enhanced gas recovery or storage. While there are few theoretical considerations on the effect of gas adsorption on permeability, a systematic laboratory investigation of the impact of gas adsorption on gas flow and permeability is still lacking. In this paper, permeability of four adsorptive gases, i.e., nitrogen, argon, methane, and carbon dioxide, was measured, along with helium permeability, for two nanoporous rock samples that have high and low total organic carbon (TOC) content, respectively. The measurements were conducted at a range of pore pressures from 150 to 1500 psi (1.03–10.34 MPa). Gas adsorption isotherms were also measured at the same conditions. A mathematical model that considers adsorption with specific boundary conditions for the experimental setup was used for data analysis. The results show that gas adsorption causes larger drop in pressure decay and greater retardation in pressure equilibrium. However, the reduction of permeability relative to helium (25%–46%) is similar for gases with different levels of adsorption, indicating the occurrence of single-layer adsorption for these gases. Comparison between the two samples further supports the concept of single-layer adsorption and signifies the impact of pore size on the permeability reduction due to adsorption. These new findings deepen the fundamental understanding and provide important clarification on the effect of gas adsorption on gas flow and permeability in nanoporous rocks.