It is essential to tackle the startup issues prior to the broad application of loop heat pipes (LHPs) in both space and terrestrial surroundings. As non-condensable gas (NCG) is an important factor affecting the startup behavior, its effects on the startup performance of an ammonia-stainless steel LHP with and without preconditioning were experimentally investigated in this work. The temperature overshoot, liquid superheat and startup time were employed as the evaluation criteria. Nitrogen was selected to simulate the NCG, which was injected into the LHP at controllable amounts. Experimental results showed that with NCG presence in the LHP, the startup could only proceed in situation 1 with preconditioning, while it could proceed in situations 1, 3 or 4 without preconditioning. For the startup in situation 1, a larger NCG inventory led to much degraded startup performance, and a higher startup heat load could benefit the startup. For the startup in situation 3, i.e. the most difficult startup situation, NCG resulted in a very high temperature overshoot during the startup, which may even exceed the maximum allowable temperature to cause a startup failure. For the startup in situation 4, the existence of NCG in the vapor grooves could facilitate the evaporation of liquid there followed by a resultant very desirable startup.