Domestic cats are popular pets, and they have personalities, with stable behavior differences between individuals. Lately, feline behavior and personality have been studied with different approaches, for example, with owner-completed questionnaires. The majority of these studies, however, lack a sufficient validation and reliability assessment of the questionnaires used. We designed an online feline behavior and personality questionnaire to collect cat behavior data from their owners. Then, we ran a factor analysis to study the structure of personality and behavior in a dataset of over 4300 cats. For validation, we studied the internal consistency, test–retest reliability, inter-rater reliability, convergent validity and discriminant validity of this questionnaire and extracted factors. In addition, we briefly examined breed differences in the seven discovered factors: fearfulness, activity/playfulness, aggression toward humans, sociability toward humans, sociability toward cats, excessive grooming and litterbox issues. Most of the rank ordering of breeds within each trait paralleled what has been found in previous studies. The validity and reliability of the questionnaire and factors were good, strengthening owner-completed questionnaires as a method to collect behavioral data from pet animals. Breed differences suggest a genetic background for personality. However, these differences should be studied further with multidimensional models, including environmental and biological variables.