Arabinoxylans (AXs) and their derived arabinoxylooligosaccharides (AXOS) are valuable functional biomolecules with promising applications in the food, brewing, and packaging industries. However, inconsistencies in extraction techniques, structural characterization, and industrial applicability remain key challenges. This review comprehensively examines various AX extraction methods from brewer's spent grain (BSG), including chemical, enzymatic, steam explosion, ultrasound-assisted, and microwave-assisted techniques, emphasizing their impact on molecular characteristics such as molecular weight, monosaccharide composition, and structural features. Furthermore, the biological activities of AX, including prebiotic, immunomodulatory, hypoglycemic, and antioxidant properties, are analyzed in relation to their structural variations. Despite their potential, knowledge gaps persist regarding how different extraction techniques influence AX bioactivity and functionality. Additionally AX is increasingly explored in synbiotic formulations, food products, beer, and biodegradable films. This review addresses these gaps by consolidating current knowledge on AX extraction, characterization, and functionality, highlighting the need for standardized methodologies and further research to optimize its industrial applications.