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Background: We investigated the association of neighborhood walkability with accelerometer‐measured physical activity (PA) and sedentary behavior (SB) and examined whether objective and subjective measures of walkability resulted in similar findings. Methods: PA and SB from the first 7689 Maastricht Study participants ages 40–75 from 2010 to 2017 were measured using accelerometers for 7 days. Mean daily step count, light‐intensity PA, moderate‐ to vigorous‐ intensity PA (MVPA), and SB were calculated. Objective walkability was measured by the 7‐component Dutch Walkability Index within 500 m Euclidean buffers around residential addresses of participants. Subjective walkability was obtained from the Abbreviated Neighborhood Environment Walkability Scale. Linear regression models analyzed the associations of walkability with PA and SB, controlling for potential confounders. Results: Objective walkability was negatively associated with light intensity PA in the most walkable quartile (b = −14.58, 95% CI = −20.94, −8.23). Compared to participants living in the least walkable neighborhoods, those in the most walkable quartile had statistically significantly higher SB levels (b = 11.64, 95% CI = 4.95, 18.32). For subjective walkability, mean daily step count was significantly higher in the most walkable quartile (b = 509.60, 95% CI = 243.38, 775.81). Higher subjective walkability was positively associated with MVPA (b = 4.40, 95% CI = 2.56, 6.23). Conclusion: Living in a neighborhood with higher objective walkability was associated with lower levels of PA and higher SB levels while higher subjective walkability was associated with higher levels of PA. These results show discordant findings and thus, the effect of walkability on participant PA and SB within our sample is to be determined. [ABSTRACT FROM AUTHOR]