Wesleyan University View Institution's Website 6 articles published in JoVE Biochemistry Förster Resonance Energy Transfer Mapping: A New Methodology to Elucidate Global Structural Features Jack Northrop1, Donald B. Oliver1,2, Ishita Mukerji1,2 1Molecular Biology and Biochemistry Department, Wesleyan University, 2Molecular Biophysics Program, Wesleyan University The study details the methodology of FRET mapping including the selection of labeling sites, choice of dyes, acquisition, and data analysis. This methodology is effective at determining binding sites, conformational changes, and dynamic motions in protein systems and is most useful if performed in conjunction with existing 3-D structural information. Developmental Biology Dissection of the Drosophila Pupal Retina for Immunohistochemistry, Western Analysis, and RNA Isolation Miles W. DeAngelis1, Ruth I. Johnson1 1Department of Biology, Wesleyan University This paper presents a surgical method for dissecting Drosophila pupal retinas along with protocols for the processing of tissue for immunohistochemistry, western analysis, and RNA-extraction. Biochemistry Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition Stacy N. Uchendu1, Angelika Rafalowski1, Erin F. Cohn1, Luke W. Davoren1, Erika A. Taylor1 1Department of Chemistry, Wesleyan University Here we present a protocol for anaerobic protein purification, anaerobic protein concentration, and subsequent kinetic characterization using an oxygen electrode system. The method is illustrated using the enzyme DesB, a dioxygenase enzyme which is more stable and active when purified and stored in an anaerobic environment. Engineering Methods for Measuring the Orientation and Rotation Rate of 3D-printed Particles in Turbulence Brendan C. Cole1, Guy G. Marcus1, Shima Parsa1, Stefan Kramel1, Rui Ni1, Greg A. Voth1 1Department of Physics, Wesleyan University We use 3D printing to fabricate anisotropic particles in the shapes of jacks, crosses, tetrads, and triads, whose alignments and rotations in turbulent fluid flow can be measured from multiple simultaneous video images. Developmental Biology Live-imaging of the Drosophila Pupal Eye Mark B. Hellerman1, Richard H. Choe1, Ruth I. Johnson1 1Biology Department, Wesleyan University This protocol presents an efficient method for imaging the live Drosophila pupal eye neuroepithelium. This method compensates for tissue movement and uneven topology, enhances visualization of cell boundaries through the use of multiple GFP-tagged junction proteins, and uses an easily-assembled imaging rig. Biology Application of Stopped-flow Kinetics Methods to Investigate the Mechanism of Action of a DNA Repair Protein F. Noah Biro1, Jie Zhai1, Christopher W. Doucette1, Manju M. Hingorani1 1Molecular Biology and Biochemistry Department, Wesleyan University Msh2-Msh6 is responsible for initiating repair of replication errors in DNA. Here we present a transient kinetics approach towards understanding how this critical protein works. The report illustrates stopped-flow experiments for measuring the coupled DNA binding and ATPase kinetics underlying Msh2-Msh6 mechanism of action in DNA repair.