The objective of this study was to examine the association between acculturation and physical activity among first generation Latina (Hispanic women) immigrants living in North Carolina. As part of the multi-site Women's Cardiovascular Health Network, 671 first generation Latina immigrants 20-50 years of age completed in-person interviews on physical activity, acculturation (measured by a language scale, length of residence in the US, and age at arrival in the US) and other potential individual and contextual correlates of physical activity. All statistical models were adjusted for age, general health, number of children in the home, marital status, and education, with self-reported physical activity as the dependent variable. Among participants, 37.4% met recommendations for physical activity, 41.9% reported insufficient activity, and 20.7% reported no moderate or vigorous activity. Latinas with higher English language acculturation were more likely to be physically active than women with lower English language acculturation. Likewise, women who arrived to the US when they were younger than 25 years were more likely to be physically active than women who arrived when they were 25 years or older. Length of residence in the US was not associated with physical activity. These relationships persisted when restricting the sample to Mexican born women. We conclude that among first generation immigrants living in North Carolina, those with higher English language acculturation or who arrived to the US at younger ages were more likely to report being physically active. These findings suggest that future epidemiologic studies and physical activity interventions should measure and tailor programs based on English-language use and age of arrival to the US among Latina immigrants.