22(S)-Hydroxycholesterol (22β-hydroxycholesterol) is a specialized oxysterol primarily used in biochemical and biomedical research to investigate cholesterol metabolism and steroid hormone biosynthesis. It acts as a key intermediate in the conversion of cholesterol to pregnenolone by the cytochrome P450 side-chain cleavage enzyme (P450scc), making it essential for studying steroidogenesis in endocrine tissues such as the adrenal glands and gonads. This compound is valuable for exploring the regulation of cholesterol homeostasis, its role in cellular signaling, and the effects of oxysterols on immune responses, inflammation, and neurodegenerative diseases. Researchers also utilize it in drug discovery efforts targeting disorders associated with dysregulated cholesterol metabolism, including cardiovascular and endocrine-related conditions.