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Two‐Hybrid Dual Bait System

Abstract Table of Contents Materials Figures Literature Cited Abstract The yeast two?hybrid system, or interaction trap, is one of the most versatile methods available with which to identify and establish

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Measurement of Oxygen Radicals and Lipid Peroxidation in Neural Tissues

Abstract Table of Contents Materials Figures Literature Cited Abstract One of the most completely validated processes involved in secondary tissue damage following acute brain or spinal cord injury and in

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Strep/FLAG Tandem Affinity Purification (SF‐TAP) to Study Protein Interactions

Abstract Table of Contents Materials Figures Literature Cited Abstract In recent years, several methods have been developed to analyze protein?protein interactions under native conditions. One of them, ta

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Metabolic Radiolabeling of Animal Cell Glycoconjugates

Abstract Table of Contents Materials Figures Literature Cited Abstract Useful information about glycoconjugates can be obtained by labeling their aglycone (noncarbohydrate) portions?e.g., labeling protein

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Nucleoside Modification with Boron Clusters and Their Metal Complexes

Abstract Table of Contents Materials Figures Literature Cited Abstract General methods for the synthesis of nucleosides modified with borane clusters and metallacarborane complexes are presented. These in

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Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation

Abstract Table of Contents Materials Figures Literature Cited Abstract Streptavidin affinity purification of protein complexes, in combination with in vivo biotinylation of critical transcription factors,

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Introduction to Atomic Force Microscopy (AFM) in Biology

Abstract Table of Contents Figures Literature Cited Abstract The atomic force microscope (AFM) has the unique capability of imaging biological samples with molecular resolution in buffer solution. In addition to providing topographical ima

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TRAF‐Mediated TNFR‐Family Signaling

Abstract Table of Contents Figures Literature Cited Abstract The tumor necrosis factor (TNF) superfamily consists of a wide variety of cell?bound and secreted proteins that regulate numerous cellular processes. In particular, TNF?family pr

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Carbohydrate‐Oligonucleotide Conjugates

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes a strategy for attaching two mannose and two galactose residues to an oligonucleotide. This conjugation can be pe

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Fluorescence Spectroscopy

Abstract Table of Contents Materials Figures Literature Cited Abstract Fluorescence is an extremely powerful tool in modern biology, physics, and chemistry laboratories. This unit focuses on methods that

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Spatiotemporal Dynamics of Kinase Signaling Visualized by Targeted Reporters

Abstract Table of Contents Materials Figures Literature Cited Abstract The advent of genetically encoded FRET?based kinase activity reporters has ushered in a new era of signal transduction research. Such

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Generation of Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) Using Directed Molecular Evolution

Abstract Table of Contents Materials Figures Literature Cited Abstract G protein?coupled receptors (GPCRs) and their signal transductions are important for both physiological and pathological processes in

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Methylation‐Sensitive Single‐Molecule Analysis of Chromatin Structure

Abstract Table of Contents Materials Figures Literature Cited Abstract Methylation?sensitive single?molecule analysis of chromatin structure is a high?resolution method for studying nucleosome positioning

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Mapping Networks of Protein‐Mediated Physical Interactions Between Chromatin Elements

Abstract Table of Contents Materials Figures Literature Cited Abstract Understanding of transcriptional regulation has advanced in recent years in part due to development of technologies which allow deter

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Chromatin Interaction Analysis Using Paired‐End Tag Sequencing

Abstract Table of Contents Materials Figures Literature Cited Abstract Chromatin Interaction Analysis using Paired?End Tag sequencing (ChIA?PET) is a technique developed for large?scale, de novo analysis

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Use of Flow Cytometric Methods to Quantify Protein‐Protein Interactions

Abstract Table of Contents Materials Figures Literature Cited Abstract A method is described for the quantitative analysis of protein?protein interactions using the flow cytometry protein interaction assa

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Construction and Use of Glycan Microarrays

Abstract Table of Contents Materials Figures Literature Cited Abstract Glycosylation is an important post?translational modification that influences many biological processes critical for development, nor

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Biosensors for Characterizing the Dynamics of Rho Family GTPases in Living Cells

Abstract Table of Contents Materials Figures Literature Cited Abstract The biosensors developed in the authors' laboratory have been based on different designs, each imparting specific strengths and weakn

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Visualization of G Protein−Coupled Receptor (GPCR) Interactions in Living Cells Using Bimolecular Fluorescence Complementation (BiFC)

Abstract Table of Contents Materials Figures Literature Cited Abstract Members of the G protein?coupled receptor (GPCR) superfamily have been shown to homo? and hetero?oligomerize both in vitro and in viv

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Installation of Site‐Specific Methylation into Histones Using Methyl Lysine Analogs

Abstract Table of Contents Materials Figures Literature Cited Abstract Chromatin structure is influenced by post?translational modifications on histones, the principal basic protein component of chromatin

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