3 articles published in JoVE
Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP) Jordan F. Hastings1, Jeremy Z.R. Han1, Robert F. Shearer1,2, Sean P. Kennedy1,3, Mary Iconomou1,4, Darren N. Saunders5, David R. Croucher1,6,7 1The Kinghorn Cancer Centre, Garvan Institute of Medical Research, 2Ubiquitin Signaling Group, Protein Signaling Program, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, 3RCSI Molecular Medicine, Royal College of Surgeons in Ireland, 4Department of Epigenetics, Max Planck Institute of Immunobiology and Epigenetics, 5School of Medical Sciences, University of New South Wales, 6St Vincent's Hospital Clinical School, University of New South Wales, 7School of Medicine and Medical Science, University College Dublin This manuscript describes the protocol for Bimolecular Complementation Affinity Purification (BiCAP). This novel method facilitates the specific isolation and downstream proteomic characterization of any two interacting proteins, while excluding un-complexed individual proteins as well as complexes formed with competing binding partners.
Preparation of an Awake Mouse for Recording Neural Responses and Injecting Tracers Michael A. Muniak1,2, Zachary M. Mayko3, David K. Ryugo2,4, Christine V. Portfors3 1Department of Neuroscience, Johns Hopkins University, 2Garvan Institute of Medical Research, 3School of Biological Sciences, Washington State University, 4Department of Otolaryngology-HNS, Johns Hopkins University Electrophysiological characterization of neuronal responses is important for understanding brain function and for guiding the placement of dyes for pathway tracing. However, many studies are performed in anesthetized animals. To understand brain function without anesthetics, we developed a method to record neuronal response properties and inject dyes in awake mouse.
DNA Methylation: Bisulphite Modification and Analysis Kate Patterson*1, Laura Molloy*1, Wenjia Qu1, Susan Clark1,2 1Epigenetics Group, Cancer Research Program, Garvan Institute of Medical Research, 2St Vincent's Clinical School, University of NSW The gold standard for DNA methylation analysis is genomic sequencing of bisulphite converted DNA. This method takes advantage of the increased sensitivity of cytosine compared with 5-methylcytosine (5-MeC) to bisulphite deamination under acidic conditions. Unmethylated cytosines can be distinguished from methylated cytosines after PCR amplification of the target genomic DNA.