Inducible Nitric Oxide Synthase Purification

Students: Alejandra Johnson, Andrew James

Faculty Mentor: Monica Lares

Chemistry

College of Science, Technology, and Business

Inducible Nitric Oxide Synthase (iNOS) plays a crucial role in immune responses by catalyzing the production of nitric oxide from L-arginine. In this project, iNOS was isolated from RAW 264.7 mouse macrophage cells using a multi-step purification process. The initial crude extraction involved mechanical homogenization and centrifugation to obtain a lysate. Subsequent purifications were performed using size-exclusion chromatography, allowing partial separation based on 10,000 kDa and 30,000 kDa sizes. We conducted a Bradford Assay and Griess Assay at each purification step to analyze total protein and our enzyme of interest to generate a purification table. We hope to show that physical purification methods are adequate without immunoaffinity techniques.