Nonreciprocal devices, such as isolators, are of great importance for optical communication and optical information processing. To bypass the limitation of a strong magnetic field imposed by the traditional Faraday magneto-optic effect, many alternative mechanisms have been proposed to demonstrate magnetic-free nonreciprocity. However, limited by the drive-induced noise, the noiseless isolator capable of working in the quantum regime has yet to be realized in the experiment. Here, we show a noiseless all-optical isolator with genuine single photons in hot atoms. We experimentally study this mechanism using an open V-type level scheme and demonstrate a low insertion loss of 0.6 dB and high isolation of 30.3 dB with bandwidth up to hundreds of megahertz. Furthermore, the nonreciprocal direction can be truly reversed only by tuning the frequency of the pump laser with the same setup. Our scheme relies on widely used optical technology and is thus universal and robust.