In battery electric buses (e-buses), the substantial energy consumption of the heating, ventilation, and air conditioning (HVAC) system can cause significant reductions of the available travel range. Additionally, HVAC systems are often operated at higher levels than what required for the thermal comfort of the passengers. Therefore, this paper proposes a method to experimentally investigate the influence of the HVAC system on the energy consumption and thermal comfort in a 12m e-bus. An appropriate thermal comfort model is identified and the required climatic input parameters are selected and measured with self-developed sensor stations. The energy consumption of the e-bus, the state of charge (SoC) of the battery and the available travel range are measured by an embedded data logger. Climatic measurements are then performed with heating on and off on a Berlin bus line in winter conditions. The results show that the energy consumption of the e-bus is increased by a factor of 1.9 with heating on, while both the SoC and travel range are reduced accordingly. Comparing the thermal comfort with heating on and off, a decrease from “comfortable” to “slightly uncomfortable but acceptable” is observed.