Used in biochemical research as a folate analog for studying folate-dependent enzymes and cellular uptake mechanisms. Acts as a probe to investigate dihydrofolate reductase (DHFR) activity and antifolate drug resistance. Employed in targeted cancer therapy studies due to its structural similarity to folic acid, enabling selective delivery to folate receptor-overexpressing cells. Also utilized in the development of enzyme inhibitors and in vitro diagnostic applications involving folate metabolism pathways.