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Phosphoproteomics by Highly Selective IMAC Protocol

Protein phosphorylation plays an important role in biological process such as cell differentiation, cell cycle control, metabolism, and apoptosis. Toward global analysis of the phosphoproteome, enrichment is an essential step to overcome analytical challenges associated w ...

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A 1D-PAGE/LC-ESI Linear Ion Trap Orbitrap MS Approach for the Analysis of Synapse Proteomes and Synaptic Protein Complexes

The 1D-PAGE/LC-ESI MS/MS approach is widely used for the qualitative and quantitative analysis of proteomes ranging from low to high complexity. As the first dimension of separation is based on SDS-PAGE, this method is compatible with the analysis of all classes of proteins including hydrop ...

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OFFGEL-Isoelectric Focusing Fractionation for the Analysis of Complex Proteome

The proteomic analysis at the peptide level is increasingly becoming a method of choice for complex samples. The success, however, depends on the development of attractive peptide fractionation methodologies to decrease the sample complexity prior to mass analysis. Recently, the OF ...

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iTRAQ-Based LC-LC MALDI TOF/TOF MS Quantitative Analysis of Membrane Proteins from Human Glioma

Various proteomic approaches are being applied in brain tumor proteomics with regard to targeted proteins of interest, to discover phenotype specific markers which could facilitate diagnosis as well as potential antitumor drug targets. iTRAQ technology is a multiplexing protein ...

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Analysis of Protein Complexes by 2D Blue Native/SDSPAGE and Antibody-Shift Assay

Blue native polyacrylamide gel electrophoresis (BN-PAGE) is a powerful strategy for the separation of native multiprotein complex (MPC). Combined with other techniques, such as SDS–PAGE, immunoblotting, mass spectrometry, and antibody-shift assay, BN-PAGE has been widely used in ...

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Enrichment of Plasma Membranes from Small Brain Samples by Aqueous Polymer Two-Phase Systems

Plasma membrane (PM) proteins take center-stage in most of the fundamental processes of the nervous system. They impart specificity to the formation of neuronal circuits, and determine the mode of neurotransmission. To accomplish these tasks, they demonstrate spatially and tempora ...

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Identification and Characterization of Protein Complexes from Total Brain and Synaptoneurosomes: Heterogeneity of Molecular Complexes in Distinct Subc

Neurons are highly polarized cells characterized by subcellular microdomains: the synapses. These compartments are specialized structures and are, for certain cellular pathways, independent from the cell body. To achieve such a functional specificity, including local mRNA tr ...

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Isolation of Synapse Subdomains by Subcellular Fractionation Using Sucrose Density Gradient Centrifugation

A protocol presents a purification of postsynaptic density (PSD), from rat brain by subcellular fractionation using solubilization of membrane with Triton X-100 and sucrose density centrifugation. The protocol also includes purification of other synapse subdomains such as syn ...

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Subcellular Fractionation of Brain Tissue Using Free-Flow Electrophoresis

Accurate annotation of protein identifications in organellar proteomics highly depends on the sample quality with special respect to contaminations from other subcellular compartments. In this respect, Free-flow electrophoresis (FFE) offers a valuable alternative to cla ...

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Dissection of Rodent Brain Regions

Dissection of brain tissue is an important step in sample preparation for (subcellular) proteomics studies. In this chapter, brain removal and separate dissection of multiple brain regions from a single brain are described in step-by-step protocol. This concerns dissection from fresh ...

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Neuroproteomics: Deciphering Brain Function and Disorders

Neuroproteomics is a branch of proteomics that specifically studies qualitatively and/or quantitatively the tissue/organelle proteomes of the nervous system. This chapter introduces the various aspects of neuroproteomics, and outlines the range of methods that are commonly ...

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Using Naturally Occurring Spinal Cord Injury in Domestic Dogs to Explore Novel Therapeutic Options

Modeling human disease in animals has traditionally been used to elucidate pathogenesis and test promising new approaches to treatment. This approach has now identified a plethora of interventions that ameliorate functional deficits associated with spinal cord injury (SCI) in ro ...

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The Cat Model of Spinal Cord Injury

The cat can recover a robust pattern of hindlimb locomotion following partial or complete spinal cord injuries. As a result, it has been instrumental in our understanding of spinal networks controlling and regulating locomotor activity. Thanks to our feline friend, spinal cord-injured ...

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Spinal Cord Injury: The Rabbit Model

This chapter gives reasons for the choice of rabbit in experimental neurobiology and includes descriptions of both the spinal cord injury models and behavioral neuroscience methods. The models provide mechanisms for hypothesis generation that can be investigated at molecular and ...

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No-Laminectomy Spinal Cord-Transected Murine Model

Most animals of contused, compressed, or transected spinal cord injury require that a laminectomy is performed. However, increasing evidence suggests that laminectomies may induce undesirable effects including neuropathic pain, spinal instabilities, longer anesthetic a ...

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Dorsal Root Injury for the Study of Spinal Cord Injury Repair

Dorsal root injury provides opportunities for highly reproducible lesions and for detailed anatomical, physiological, and behavioral outcome assessment with high precision and validity. Dorsal root injury models are used to several aspects of relevance to spinal cord injury re ...

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Spinal Cord Injury: The Lamprey Model

Following severe spinal cord injury (SCI), brain locomotor command systems are disconnected from spinal motor networks, resulting in paralysis caudal to the lesion. In higher vertebrates, such as birds and mammals, the CNS is a restrictive environment for axonal regeneration. As a result, ...

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Acute Spinal Cord Injury In Vitro: Insight into Basic Mechanisms

Spinal injury is produced by traumatic and nontraumatic causes that often induce long-term disability. In order to limit tissue damage and functional impairment, it is important to intervene at an early stage after the primary insult to restrain damage extension. This objective is diffic ...

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The Pathological Findings in Traumatic Injury to the Human Spinal Cord

The anatomical pattern of damage following upon traumatic injury to the human spinal cord is dependent upon the manner in which the injury is sustained. Hyperflexion typically results in anterior spinal cord injury, hyperextension in central spinal cord injury, stab wounds in hemisect ...

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The Spinal Cord: Functional Organization, Diseases, and Dysfunctions

The spinal cord constitutes a key structure of the central nervous system. It is involved in the transmission of signals between the brain and the rest of the body through numerous ascending and descending pathways. It also contains relatively simple reflex arcs as well as more complex neuronal c ...

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