The gas-flow reduction factor of the second forward Differential Pumping Section (DPS2-F) for the KATRIN experiment was determined using a dedicated vacuum-measurement setup and by detailed molecular-flow simulation of the DPS2-F beam tube and of the measurement apparatus. In the measurement, non-radioactive test gases deuterium, helium, neon, argon and krypton were used, the input gas flow was provided by a commercial mass-flow controller, and the output flow was measured using a residual gas analyzer, in order to distinguish it from the outgassing background. The measured reduction factor with the empty beam tube at room temperature for gases with mass 4 is 1.8(4) × 10 4 , which is in excellent agreement with the simulated value of 1.6 × 10 4 . The simulated reduction factor for tritium, based on the interpolated value for the capture factor at the turbo-molecular pump inlet flange is 2.5 × 10 4 . The difference with respect to the design value of 1 × 10 5 is due to the modifications in the beam tube geometry since the initial design, and can be partly recovered by reduction of the effective beam tube diameter.