Max Planck Institute of Immunobiology and Epigenetics 3 articles published in JoVE Immunology and Infection Targeted Metabolomics on Rare Primary Cells Katharina M. Glaser*1, Mirijam Egg*1, Sebastian Hobitz1, Michael Mitterer1, Dominik Schain-Zota1, Katharina Schönberger1, Konrad Schuldes1, Nina Cabezas-Wallscheid1, Tim Lämmermann1, Angelika Rambold1, Joerg M. Buescher1 1Max Planck Institute of Immunobiology and Epigenetics Here, we present a protocol to accurately and reliably measure metabolites in rare cell types. Technical improvements, including a modified sheath fluid for cell sorting and the generation of relevant blank samples, enable a comprehensive quantification of metabolites with an input of only 5000 cells per sample. Biology Specific Labeling of Mitochondrial Nucleoids for Time-lapse Structured Illumination Microscopy Visnja Jevtic1, Petra Kindle1, Sergiy V. Avilov1 1Imaging Facility, Max Planck Institute of Immunobiology and Epigenetics The protocol describes specific labeling of mitochondrial nucleoids with a commercially available DNA gel stain, acquisition of time lapse series of live labeled cells by super-resolution structured illumination microscopy (SR-SIM), and automatic tracking of nucleoid motion. Immunology and Infection 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.
