Van Andel Research Institute View Institution's Website 6 articles published in JoVE Cancer Research Intraductal Delivery and X-ray Visualization of Ethanol-Based Ablative Solution for Prevention and Local Treatment of Breast Cancer in Mouse Models Elizabeth Kenyon1,2, Erin K. Zaluzec1,3, Katherine Powell1,2, Maximilian Volk1,4, Shatadru Chakravarty2,5, Jeremy Hix2,6, Ripla Arora7,8, Jennifer J. Westerhuis9,10, Matti Kiupel11, Erik M. Shapiro2,6, Lorenzo F. Sempere1,2 1Precision Health Program, Michigan State University, 2Department of Radiology, College of Human Medicine, Michigan State University, 3Department of Pharmacology & Toxicology, College of Veterinary Medicine, Michigan State University, 4College of Osteopathic Medicine, Michigan State University, 5Advanced Materials Characterization Laboratory/Materials Research Center, Missouri University of Science and Technology, 6Institute for Quantitative (IQ) Health Science and Engineering Advanced Molecular Imaging Facility, Michigan State University, 7Department of Obstetrics Gynecology and Reproductive Biology, College of Human Medicine, Michigan State University, 8Institute for Quantitative (IQ) Health Science and Engineering, Michigan State University, 9Van Andel Research Institute, 10Miltenyi Biotec, 11Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Michigan State University A method of intraductal injection of reagents for an ethanol-based ablative solution to the mouse mammary ductal tree for in vivo imaging and breast cancer prevention is described. Injection directly into the nipple opening allows for targeting mammary epithelial cells with minimal collateral tissue damage. Genetics In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression Nicole A. Vander Schaaf1, Shirley Oghamian2, Jin-A Park1, Liang Kang1, Peter W. Laird1, Kwang-Ho Lee1 1Center for Epigenetics, Van Andel Research Institute, 2Amgen This protocol outlines the steps needed to generate a model system in which the transcription of an endogenous gene of interest can be conditionally controlled in live animals or cells using enhanced lac repressor and/or tet activator systems. Biochemistry Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy Emery Haley*1, Wooyoung Choi*1, Chen Fan1, Weinan Sun2,3, Juan Du1, Wei Lü1 1Van Andel Institute, 2Vollum Institute, 3Janelia Research Campus This protocol describes techniques used to determine ion channel structures by cryo-electron microscopy, including a baculovirus system used to efficiently express genes in mammalian cells with minimum effort and toxicity, protein extraction, purification, and quality checking, sample grid preparation and screening, as well as data collection and processing. Biochemistry Analysis of Histone Antibody Specificity with Peptide Microarrays Evan M. Cornett1, Bradley M. Dickson1, Scott B. Rothbart1 1Center for Epigenetics, Van Andel Research Institute This manuscript describes methods for applying peptide microarray technology to specificity profiling of antibodies that recognize histones and their post-translational modifications. Biochemistry Multimer-PAGE: A Method for Capturing and Resolving Protein Complexes in Biological Samples Tyler Rhinesmith1, Bryan A. Killinger2, Akhil Sharma3, Anna Moszczynska3 1Physiology, Michigan State University, 2Center for Neurodegenerative Science Van Andel Institute, 3Pharmaceutical Sciences, Wayne State University A method for stabilizing and separating native protein complexes from unmodified tissue lysate using an amine-reactive protein cross-linker coupled to a novel two-dimensional polyacrylamide gel electrophoresis (PAGE) system is presented. Biology Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples Chen Lu1, Joshua L. Wonsidler1, Jianwei Li2, Yanming Du1, Timothy Block3, Brian Haab4, Songming Chen1 1Institute for Hepatitis and Virus Research, 2Department of Microbiology and Immunology, Thomas Jefferson University, 3Drexel University College of Medicine, 4Van Andel Research Institute In this study, we describe an improved protocol for a multiplexed high-throughput antibody microarray with lectin detection method that can be used in glycosylation profiling of specific proteins. This protocol features new reliable reagents and significantly reduces the time, cost, and lab equipment requirements as compared to the previous procedure.