Isotope analysis has become an increasingly valuable tool in forensic anthropology casework over the past decade. Modern-day isotopic investigations on human remains have integrated the use of multi-isotope profiles (e.g. C, N, O, H, S, Sr, and Pb) as well as isotopic landscapes (“isoscapes”) from multiple body tissues (e.g. teeth, bone, hair, and nails) to predict possible region-of-origin of unidentified human remains. Together, data from various isotope analyses provide additional lines of evidence for human identification, including a decedent’s possible region-of-birth, long-term adult residence, recent travel history, and dietary choices. Here, we present the basic principles of isotope analysis and provide a brief overview of instrumentation, analytical standards, sample selection, and sample quality measures. Finally, we present case studies that reflect the diverse applications of isotope analysis to the medicolegal system before describing some future research directions. As shown herein, isotope analysis is a flexible and powerful geolocation tool that can provide new investigative leads for unidentified human remains cases.