Genentech, Inc. 9 articles published in JoVE Medicine Synthetic Antigen Controls for Immunohistochemistry Charles A. Havnar1, Kathy J. Hötzel1, Charles A. Jones1, Carmina M. Espiritu1, Linda K. Rangell1, Franklin V. Peale1 1Research Pathology, Genentech, Inc. This work documents a simple method to create synthetic antigen controls for immunohistochemistry. The technique is adaptable to a variety of antigens in a wide range of concentrations. The samples provide a reference with which to assess intra- and inter-assay performance and reproducibility. Biochemistry gP2S, an Information Management System for CryoEM Experiments Dorota Wypych1, Daniel Kierecki1, Filip M. Golebiowski1, Alexis Rohou2 1Roche Polska Sp. z o.o., 2Department of Structural Biology, Genentech gP2S is a web application for the tracking of cryoEM experiments. Its main features are described, as are the steps required to install and configure the application. Once configured, the application allows one to accurately record metadata associated with negative stain and cryoEM experiments. Cancer Research Automated Dissection Protocol for Tumor Enrichment in Low Tumor Content Tissues Charles A. Havnar1, Oliver Zill2, Jeff Eastham1, Jeffrey Hung1, Manana Javey3, Emmanuel Naouri3, Jennifer Giltnane1, Justin M. Balko4, Andrew Wallace2, Nicolas Lounsbury2, Daniel Oreper2, Sarajane Saturnio1, G-Y Yang5, Amy A. Lo1 1Departments of Research Pathology, Genentech, 2Bioinformatics & Computational Biology, Genentech, 3Roche Sequencing Solutions, Hacienda Drive, 4Department of Medicine, Vanderbilt University Medical Center, Medical Center Drive, 5Department of Pathology, Northwestern University, Feinberg School of Medicine Digital annotation with automated tissue dissection provides an innovative approach to enriching tumor in low tumor content cases and is adaptable to both paraffin and frozen tissue types. The described workflow improves accuracy, reproducibility and throughput and could be applied to both research and clinical settings. Biochemistry Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions Bushra Husain1, Sairupa Paduchuri1, Sree R. Ramani2, Nadia Martinez-Martin1 1Receptor Discovery group, Microchemistry, Proteomics and Lipidomics Department, Genentech, 2Portfolio Management and Operations, Genentech Here, we present a protocol to screen extracellular protein microarrays for identification of novel receptor-ligand interactions in high throughput. We also describe a method to enhance detection of transient protein-protein interactions by using protein-microbead complexes. Immunology and Infection Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry Ryan M. Baxter*1, Daniel S. Kong*1, Josselyn E. Garcia-Perez1, William E. O'Gorman2, Elena W.Y. Hsieh1,3 1Department of Immunology and Microbiology, University of Colorado School of Medicine, 2OMNI Biomarkers, Development Sciences, Genentech, 3Department of Pediatrics, Division of Allergy and Immunology, University of Colorado School of Medicine Here we describe a single-cell proteomic approach to evaluate immune phenotypic and functional (intracellular cytokine induction) alterations in peripheral whole blood samples, analyzed via mass cytometry. Immunology and Infection Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice Anna Zychlinsky Scharff1, Matthew L. Albert1,2, Molly A. Ingersoll1 1Unité d’Immunobiologie des Cellules Dendritiques, Department of Immunology, Institut Pasteur, INSERM U818, 2Genentech The established model of transurethral catheterization of mice allows the study of bladder pathologies, including urinary tract infection, but can only be performed in females. A new model of male transurethral instillation, presented here, will enable research in an area marked by strong clinical and epidemiological differences between the sexes. Chemistry Application and Methodology of the Non-destructive 19F Time-domain NMR Technique to Measure the Content in Fluorine-containing Drug Products Maria Victoria Silva Elipe1, Lan Li1, Karthik Nagapudi2, Alan M. Kook3, Carlos Cobas4, Isaac Iglesias4, Chen Peng4 1Department of Attribute Sciences, Amgen, Inc., 2Small Molecule Pharmaceutical Sciences, Genentech, Inc., 3NMR Service + Consulting, 4Mestrelab Research A simple and non-destructive technique that measures the average content of drug substances in formulated drug products containing fluorine using low-field fluorine-19 (19F) time-domain (TD) nuclear magnetic resonance (NMR) is presented here. The technique can be applied to the development and manufacturing of drugs in the pharmaceutical industry. Medicine Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia Yu Ouyang1, Martin S. Judenhofer1, Jeffrey H. Walton1,2, Jan Marik3, Simon P. Williams3, Simon R. Cherry1,4 1Department of Biomedical Engineering, University of California, Davis, 2Nuclear Magnetic Resonance Facility, University of California, Davis, 3Biomedical Imaging, Genentech, Inc, 4Department of Radiology, University of California, Davis The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired. Neuroscience Imaging Cleared Intact Biological Systems at a Cellular Level by 3DISCO Ali Ertürk1, Daniel Lafkas2, Cecile Chalouni3 1Neuroscience, Genentech, Inc., 2Department of Discovery Oncology, Genentech, Inc., 3Department of Pathology, Genentech, Inc. To obtain high-resolution images of fluorescently labeled cells within large tissues, ideally, the biological samples should be imaged without sectioning. 3DISCO is a straightforward tissue clearing procedure based on sequential incubation with organic solvents. Upon clearing, the organs become transparent allowing an end-to-end laser scan of the specimen.
