Previous research has identified associations between work-family conflict (WFC) and health outcomes (e.g., musculoskeletal pain). This study investigated whether WFC and family-work conflict explain relationships between exposure to work-related hazards and musculoskeletal pain and stress for workers undertaking some or all of their work at home. Possible differences by home workspace location were also explored. Longitudinal survey data were collected from workers in Australia engaged in work from home for at least two days per week. Data was collected at four timepoints approximately 6 months apart (Baseline [October 2020] n = 897; Wave 1 [May/June 2021] n = 368; Wave 2 [October/November 2021] n = 336; Wave 3 [May 2022] n = 269). Subjective measures of work-related psychosocial hazards, occupational sitting and physical activity, musculoskeletal pain, and stress were collected via an online questionnaire. Mediation analyses were conducted using the R package "mediation." Analyses were also conducted with the data set stratified by home office location, using R version 4.1.3. Both WFC and family-work conflict acted as mediators between psychosocial work-related hazards and musculoskeletal pain and stress. WFC mediated more relationships than family-work conflict. Location of home workspace was important, particularly for those working in a space at home where they may be subject to interruptions. Addressing WFC is a legitimate means through which musculoskeletal pain and stress can be reduced. Organizational risk management strategies need to address all work-related risks, including those stemming from work-life interaction.