Chronic stress is both theoretically and methodologically challenging to operationalize through biomarkers. Yet minimally invasive, field-friendly biomarkers of chronic stress are valuable in research linking biology and culture, seeking to understand differential patterns of human development across ecological contexts, and exploring the evolution of human sociality. For human biologists, a central question in measurement and interpretation of biomarkers is how stress-responsive physiological systems are regulated across diverse human ecologies. This article aims to describe a conditional toolkit for human biologists interested in the study of chronic stress, highlighting a mix of longstanding and novel biomarkers, with special focus on hair/fingernail cortisol, latent herpesvirus antibodies, allostatic load indices, and serial/ambulatory data collection approaches. Future trends in chronic stress biomarker research, including epigenetic approaches, are briefly considered. This overview considers: (1) challenges in separating a distinctly psychosocial dimension of chronic stress from adversity more broadly; (2) essential characteristics of human ecology that shape interpretation; (3) retrospective vs. longitudinal sampling; (4) the role of age, developmental effects, and local biologies; (5) different timescales of chronicity; and (6) the role of culture.