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Max Planck Institute of Immunobiology and Epigenetics

4 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)
Jordan F. Hastings 1, Jeremy Z.R. Han 1, Robert F. Shearer 1,2, Sean P. Kennedy 1,3, Mary Iconomou 1,4, Darren N. Saunders 5, David R. Croucher 1,6,7
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.

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Biology

Specific Labeling of Mitochondrial Nucleoids for Time-lapse Structured Illumination Microscopy
Visnja Jevtic 1, Petra Kindle 1, Sergiy V. Avilov 1
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.

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Genetics

Isolation and Processing of Murine White Adipocytes for Transcriptome and Epigenome Analyses
Chih-Hsiang Yang 1,2, John Longinotto 2, Ilaria Panzeri 1,2, Laura Arrigoni 2, Vanessa Wegert 1, Steffen Heyne 2, Gabriel Seifert 3, Adelheid Lempradl 1,2, Ulrike Bönisch 2, John Andrew Pospisilik 1,2
1Van Andel Institute, 2Max Planck Institute of Immunobiology and Epigenetics, 3Department of General and Visceral Surgery, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg

The present protocol summarizes a universal method for isolating, purifying, and upstream processing of murine white adipocytes optimized for downstream total RNA sequencing, Nuclei Extraction by SONication (NEXSON), and ChIP-seq.

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Immunology and Infection

Targeted Metabolomics on Rare Primary Cells
Katharina M. Glaser *1, Mirijam Egg *1, Sebastian Hobitz 1, Michael Mitterer 1, Dominik Schain-Zota 1, Katharina Schönberger 1, Konrad Schuldes 1, Nina Cabezas-Wallscheid 1, Tim Lämmermann 1, Angelika Rambold 1, Joerg M. Buescher 1
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.

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