Stress is a major influence on the quality of life in our fast-moving society. This paper describes a standardized and contemporary protocol that is capable of inducing moderate psychological stress in a laboratory setting. Furthermore, it evaluates its effects on physiological biomarkers. The protocol called “THM-Stresstest” mainly consists of a rest period (30 min), an app-based stress test under the surveillance of an audience (4 min) and a regeneration period (32 min). We investigated 12 subjects to evaluate the developed protocol. We could show significant changes in heart rate variability, electromyography, electro dermal activity and salivary cortisol and α-amylase. From this data we conclude that the THM-Stresstest can serve as a psychobiological tool for provoking responses in the cardiovascular-, the endocrine and exocrine system as well as the sympathetic part of the central nervous system.