The biopsychosocial model of challenge and threat (BPS-CT) is a powerful framework linking psychological processes to reliable patterns of cardiovascular responses during motivated performance situations. Specifically, the BPS-CT poses challenge and threat as two motivational states that can emerge in response to a demanding, self-relevant task, where greater challenge arises when perceived resources are higher than demands, and greater threat arises when perceived resources are lower than demands. By identifying unique patterns of physiological responses associated with challenge and threat, respectively, the BPS-CT affords insight into subjective appraisals of resources and demands, and their determinants, during motivated performance situations. Despite its broad utility, lack of familiarity with physiological concepts and difficulty with identifying clear guidelines in the literature are barriers to wider uptake of this approach by behavioral researchers. Our goal is to remove these barriers by providing a comprehensive, step-by-step tutorial on conducting an experiment using the challenge and threat model, offering concrete recommendations for those who are new to the method, and serving as a centralized collection of resources for those looking to deepen their understanding. The tutorial spans five parts, covering theoretical introduction, lab setup, data collection, data analysis, and appendices offering additional details about data analysis and equipment. With this, we aim to make challenge and threat research, and the insights it offers, more accessible to researchers throughout the behavioral sciences.