This fully updated edition explores methods involving sphingosine-1-phosphate (S1P), a bioactive lysophospholipid which has become the focus of much research interest as it has widespread developmental and physio-pathological actions, controlling events within the nervous, reproductive, gastrointestinal, vascular, respiratory, and immune systems, in addition to having a prominent role in cancer, early mammalian embryogenesis, and stem cells. Here, worldwide experts in the S1P field describe in-depth techniques in an array of cell types and with various physiological applications, showcasing the important effects of S1P in development and in physiopathology. As a volume in the Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the relevant materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls. Authoritative and timely, Sphingosine-1-Phosphate: Methods and Protocols, Second Edition is another key resource for scientists working in this fast-moving and dynamic field.