Gambling has typically been considered a predominately male activity. However, recent prevalence surveys have shown greater numbers of females are now gambling. Much of the gambling literature suggests online gamblers are more likely to be male, and that problem gamblers are more likely to be male. Males and females are also likely to be gambling for different reasons and have a preference for different gambling activities. Little is known about the pattern of play among female online gamblers. The aim of this survey was to develop a better profile of female online gamblers and to examine any gender differences between males and females in terms of how and why they gamble online, their frequency of online gambling, patterns of play, as well as attitudes to online gambling. The survey was posted on 32 international online gambling websites and was completed by 975 online gamblers (including 175 female online gamblers). Chi-square tests of association were conducted to examine the association between gender and a range of variables. The results showed that females had been gambling online for a shorter duration of time than males, had much shorter online gambling sessions, different motivations for gambling online (i.e., to practice for free, to spend less money and out of boredom), and experienced online gambling differently to males, with increased feelings of guilt and shame for gambling online. This suggests there is still a stigma around gambling particularly evident among females in this study. The findings indicate that clinicians and treatment providers need to be aware of these potential gender differences in online gambling to develop appropriately tailored interventions.