In the past few years, the application of proteomics to examine the molecular mechanisms underlying (mal-)functioning of the nervous system and brain disorders has risen steeply, which in many cases has yielded novel insights. As neuroproteomics is a new research field involving the use of a number of high-end analytical instruments and technologies, both promises and pitfalls may not be well appreciated by the researchers, and the design of a proper proteomics experiment may not be obvious. Neuroproteomics brings together a collection of detailed protocols written by experienced professionals in the field in order to cover aspects of various dimensions of this new technology, together with some established methods that have supporting roles in the workflow of neuroproteomics. Written for the popular Neuromethods series, the chapters in this book contain step-by-step, easy to follow instruction along with practical tips and key implementation advice essential for successful implementation of the techniques.

Authoritative and cutting-edge, Neuroproteomics will be of great assistance to all researchers seeking to design and execute their own proteomics experiments in an optimal way and continue moving forward this exciting area of study.

Covers both techniques involving cutting-edge technology and more established methods Features clear, practical protocols for readily reproducible results Serves as an ideal guide to the designing of successful neuroproteomics experiments Includes supplementary material: sn.pub/extras

Inhalt

Neuroproteomics: Deciphering Brain Function and Disorders.- Dissection of Rodent Brain Regions.- Subcellular Fractionation of Brain Tissue Using Free Flow Electrophoresis.- Isolation of Synapse Sub-Domains by Subcellular Fractionation Using Sucrose Density Gradient Centrifugation.- Enrichment of Plasma Membranes from Small Brain Samples by Aqueous Polymer Two-Phase Systems.- Identification and Characterization of Protein Complexes from Total Brain and Synaptoneurosomes: Heterogeneity of Molecular Complexes in Distinct Subcellular Domains.- Analysis of Protein Complexes by 2D Blue Native/SDS-PAGE and Antibody-Shift Assay.- 2-Dimensional Gel Electrophoresis-Based Proteomic Analysis of Brain Synapses.- 2-Dimensional BAC-SDS Polyacrylamide Gel Electrophoresis for the Fractionation and Identification of Synaptic Vesicle Proteins and the Presynaptic Active Zone.- iTRAQ-Based LC-LC MALDI TOF/TOF MS Quantitative Analysis of Membrane Proteins from Human Glioma.- OFFGEL-Isoelectric Focusing Fractionation for the Analysis of Complex Proteome.- A 1D-PAGE/LC-ESI Linear Ion Trap Orbitrap MS Approach for the Analysis of Synapse Proteomes and Synaptic Protein Complexes.- SDS-PAGE Immuno-Blot Analysis.- Phosphoproteomics by Highly Selective IMAC Protocol.- Global Analysis of Ubiquitination.- High-Throughput High-Content Functional Image Analysis of Neuronal Proteins Implicated in Parkinson's Disease.- Neuropeptidomics of the Mammalian Brain.- In-Depth Analysis of the Cerebrospinal Fluid Proteome and Biomarker Discovery: Abundant Protein Depletion Sample Pretreatment Method.- Lipidomics of the Nervous System: Phospholipidomics by Liquid Chromatography Coupled to Mass Spectrometry or Tandem Mass Spectrometry.- BioinformaticsProcedures for Analysis of Quantitative Proteomics Experiments Using iTRAQ.- Statistical Analysis of Spectral Count Data Generated by Label-Free Tandem Mass Spectrometry-Based Proteomics.