Sedentary behavior (SB) is a major contributor to obesity and significant morbidity and mortality in adolescence and adulthood, yet measurement of SB is still evolving. The purpose of this study was to assess the degree of construct validity of the inclinometer function and single-axis and vector magnitude accelerometry metrics of the ActiGraph GT3X+ in objectively measuring SB and physical activity in 28 young adult university students who performed nine semi-structured activities, each for five minutes: lying, sitting, reading, seated video gaming, video watching, seated conversation, standing, stationary biking, and treadmill walking. Inclinometry and four output metrics from the ActiGraph were analyzed in comparison to direct observation by a researcher recorded each minute. For overall accuracy in measuring both SB and physical activity, all four accelerometer metrics (94.7–97.8%) outperformed the inclinometer function (70.9%). Vector magnitude accelerometry with a threshold of 150 counts per minute as the cut point for sedentary behavior was superior to other methods. While accelerometry was more accurate overall at detecting the behaviors tested, inclinometry had some advantages over accelerometry methods at detecting walking, biking, and standing. The findings support use of accelerometry as a valid objective measure of body movement, while use of inclinometry as a sole measure is not recommended. Additional research would be beneficial to improve the calibration of the inclinometer and explore ways of combining this with accelerometer data for objectively measuring SB and physical activity. © 2015 Wiley Periodicals, Inc.