The road tarantula Eupalaestrus weijenberghi shows a strongly female-biased sex ratio since adult females live several years while adult males live only for 2 months. In this scenario selective males could be expected. However, several factors such as the rates of reproduction of each sex, degree of sexual selectivity and synchronicity of female receptiveness determine the operational sexual ratio and mating system of the species. Our objective was to determine the mating rates and mating tactics for females and males of E. weijenberghi and their variation throughout the reproductive period. Four hundred sexual encounters among 20 females and 20 males in all possible pair-wise combinations were carried out during 29 days, a brief but intense experimental period, as it also occurs in the field. Mating success differed strongly between sexes. Females mated once: five females mated at the first attempt, eight initially rejected males and copulated in subsequent attempts. Half of the males did not copulate and the others copulated 1-3 times. Mated females actively rejected males. Results indicate a mating system with monogamous females and polygamous males. Not all the females were receptive in every reproductive season. We suggest that female monogamy drastically affects the operational sex ratio, since several females were unreceptive after a single copulation, directly diminishing the male potential reproductive rate. This is the first experimental approach to estimate tarantula mating systems, their determinants and the consequences of the strategies shown by each sex.