The association between the subjective experience of pain-related disability (PRD) and device-measured physical activity (PA) and sedentary behavior (SB) in overweight and obese adults is not well known. The aim of this study was to investigate the associations of pain markers with accelerometer-measured SB duration and different intensities of PA among physically inactive middle-aged adults with overweight or obesity. This cross-sectional analysis included 72 subjects (27 men) with mean age of 57.9 (SD 6.7) years and mean BMI of 31.6 (SD 4.1) kg/m2. SB and standing time (ST), breaks in sedentary time, light physical activity (LPA) and moderate-to-vigorous physical activity (MVPA) were measured for four consecutive weeks (mean 25 days, SD 4) with a hip-worn triaxial accelerometer. Headache, musculoskeletal pain, back pain, and PRD were assessed by visual analog scales (VAS) and using the Oswestry disability index (ODI). RAND-36 questionnaire was applied to assess health-related quality of life. The associations were studied by linear models. ST was positively and SB proportion was negatively associated with PRD when adjusted for age, sex, BMI, accelerometry duration, MVPA, pain medication use, and general health perceptions assessed by RAND-36. No associations were found between ST and back pain. SB or different PA intensities were not associated with pain experience at specific sites. Longer daily ST, but not LPA or MVPA is associated with higher level of PRD. Correspondingly, higher proportion of SB is associated with lower level of PRD. This suggests that individuals with PRD prefer to stand, possibly to cope with pain. These results may highlight the importance of habitual standing behaviors in coping with experienced PRD in adults with overweight or obesity. © 2021 Walter de Gruyter GmbH, Berlin/Boston.