Oats (Avena sativa L.) are an important cereal crop with diverse applications in both food and forage. Oat β-glucan has gained attention for its beneficial biological activities, such as reducing cardiovascular risk, preventing diabetes, and enhancing intestinal health. Despite its potential, more comprehensive research is required to explore its preparation, modification, bioactivities, and applications. This review highlights recent advancements in the determination and preparation of oat β-glucan, explores its biological activities and mechanisms, and examines the impact of food processing techniques on its properties. This review is intended to provide a theoretical foundation and reference for the development and application of oat β-glucan in the functional food industry.