Physical Sciences and Mathematics Commons^™

Complementary Spectroscopic Assays For Investigating Protein−Ligand Binding Activity: A Project For The Advanced Chemistry Laboratory, David Mascotti, Mark Waner

Mark J. Waner

A protein−ligand binding, guided-inquiry laboratory project with potential application across the advanced undergraduate curriculum is described. At the heart of the project are fluorescence and spectrophotometric assays utilizing biotin-4-fluorescein and streptavidin. The use of the same stock solutions for an assay that may be examined by two distinct spectroscopic techniques offers an opportunity to discuss not only protein−ligand binding but also to compare instrumental techniques and to discuss the underlying physical principles that are the origins of the assays. In addition to critically evaluating analytical techniques, students are pushed to develop quantitative pipetting, estimation, and experimental-design skills to collect and …