1Neuroscience Center of Excellence, Louisiana State University Health Sciences Center
Drosophila is famous for its powerful genetic manipulation, but not for its suitability of in-depth biochemical analysis. Here we present a TAP-based procedure to identify interacting partners of any protein of interest from the fly brain. This procedure can potentially lead to new avenues of research.
JoVE Immunology and Infection
1INSERM UMR 1043, CNRS UMR 5282, Université Toulouse 3, 2UMR Viroligie, INRA ENVA ANSES
These past 15 years, canine adenovirus type 2 (CAV2)-derived vectors have proven their efficiency to transduce cells in vitro and in vivo and are widely used for vaccination and gene therapy. Here, we describe a procedure to construct, produce and purify CAV2 vectors, giving rise to high-titer viral suspensions.
1Department of Physiology, Medical College of Wisconsin
This article describes an in situ hybridization protocol optimized for colormetric detection of microRNA expression in formalin fixed kidney sections.
1Institute for Research on Cancer and Ageing of Nice, UMR CNRS 7284 - INSERM, U1081 - UNS, Centre Antoine Lacassagne, University of Nice - Sophia Antipolis
We describe here an accurate, reproducible and convenient biochemical method for titration of glycogen in vitro. This technique uses the Abcam Glycogen assay kit and is based on successive hydrolysis of glycogen to glucose and glucose titration by fluorescence.
1School of Molecular Biosciences, Washington State University, 2Center for Reproductive Biology, Washington State University
<em>Caenorhabditis elegan</em> is a useful model to explore the functions of polyunsaturated fatty acids in development and physiology. This protocol describes an efficient method of supplementing the <em>C. elegans</em> diet with polyunsaturated fatty acids.
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), University of Heidelberg
Protein conformation and dynamics are key to understanding the relationship between protein structure and function. Hydrogen exchange coupled with high-resolution mass spectrometry is a versatile method to study the conformational dynamics of proteins as well as characterizing protein-ligand and protein-protein interactions, including contact interfaces and allosteric effects.
1Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, 2IKERBASQUE, Basque Foundation for Science
Drosophila S2 cells and cultured neurons are great systems for imaging of motor-driven organelle transport in vivo. Here we describe detailed protocols for culturing both cell types, their imaging and analysis of transport.
JoVE Immunology and Infection
1CNRS UMR 7278, IRD198, INSERM U1095, Aix-Marseille Université, 2Department of Molecular Medicine and Medical Biotechnology, University of Naples "Federico II"
Macrophages have long been recognized as a critical component of the innate and adaptive immune responses. The recent explosion of knowledge pertaining to evolutionary, genetic, and biochemical aspects of the interaction between macrophages and microbes has renewed scientific attention to macrophages. This article describes a method to differentiate macrophages from mouse bone marrow.
JoVE Applied Physics
1Department of Physics and Astronomy, University College London, 2CERN, 3Physics Division, Lawrence Berkeley National Laboratories
This paper demonstrates a protocol for recasting experimental simplified model limits into conservative and aggressive limits on an arbitrary new physics model. Publicly available LHC experimental results can be recast in this manner into limits on almost any new physics model with a supersymmetry-like signature.
1Department of Chemical Engineering, Massachusetts Institute of Technology, 2David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
Rapid mechanical deformation of cells has emerged as a promising, vector-free method for intracellular delivery of macromolecules and nanomaterials. This protocol provides detailed steps on how to use the system for a broad range of applications.