Biomembrane Protocols

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ISBN: 9780896032507 | 0896032507
Cover: Paperback
Copyright: 6/1/1994

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This second volume of protocols offers the most comprehensive set of modern analytical techniques available for studying the architecture and function of membranes. It features the application of biochemical, spectroscopic, and fluorimetric methods to the investigation of molecular topology, the dynamic aspects of membrane structure, and ion transport. Antibody technology, analysis of molecules involved in intracellular signaling, and receptor-ligand interactions are also covered.

Crystallization of Membrane Proteins for X-Ray Analysis
Determination of Cell Surface Polarity by Solid-Phase Lactoperoxidase Iodination
Biochemical Methods to Determine Cell Surface Topography
Labeling of Oxidized Glycoproteins with 3H-Borohydride
Identification and Separation of Integral Membrane Proteins Using Triton X-114
Surface-Selective Labeling of Polarized Proteins on Tight Epithelia Using Sulfo-N-Hydroxysuccinimido-Biotin (S-NHS-B
Use of Antipeptide Antibodies for the Isolation and Study of Membrane Proteins
Preparation of Antibodies
Affinity-Purification of Antipeptide Antibodies
Purification of Membrane Proteins by Immunoaffinity Chromatography
Competitive ELISA for Determining Membrane Protein Topology
The Production of Monoclonal Antibodies to Membrane Proteins
Production of Hybridoma Cell Lines
Screening for Monoclonal Antibody Production
Purification of a Membrane Protein (Ca2+/Mg2+-ATPase) and Its Reconstitution into Lipid Vesicles
Measurement of Protein-Protein Interactions in Reconstituted Membrane Vesicles Using Fluorescence Spectroscopy
Measurement of Lipid-Protein Interactions in Reconstituted Membrane Vesicles Using Fluorescence Spectroscopy
Determination of the Transverse Topography of Membrane Lipids Using Enzymes and Covalent Labels as Probes
Determination of the Transverse Topography of Membrane Phospholipids Using Phospholipid Transfer Proteins as Tools
Prothrombinase Complex as a Tool to Assess Changes in Membrane Phospholipid Asymmetry
Fluorescent Glycerolipid Probes: Synthesis and Use for Examining Intracellular Lipid Trafficking
Synthesis of Fluorescent Glycerolipid Probes
Delivery of Fluorescent Glycerophospholipid Probes to Cultured Cells
Synthesis and Use of Spin-Labeled Lipids for Studies of the Transmembrane Movement of Phospholipids
Measurement of Membrane Fluidity and Membrane Fusion with Fluorescent Probes Extraction and Assay of Cyclic Nucleotides
Analysis of G-Proteins Regulating Signal Transduction Pathways
Identification of G-Proteins
GTPase Studies
Reconstitution of Second Messenger Pathways in Permeabilized Cell Preparations Using GTP Analogs and Receptor Ligands
Assay of Protein Kinases and Protein Phosphorylation
Protein Kinase-Mediated Phosphorylation Events
Measurement of Protein Kinase Expression
Measurement of Protein Kinase Activity and Translocation
Analysis of Cellular Phosphoinositides and Phosphoinositols: Extraction and Simple Analytical Procedures
Biosynthesis and Extraction of Phosphoinositides and Phosphoinositols
Separation of Phosphoinositols by Anion-Exchange Chromatography
Separation of Phosphoinositides by Thin-Layer Chromatography
Deacylation of Phospholipids and Separation of Products by Anion-Exchange Chromatography
Analysis of Cellular Phosphoinositides and Phosphoinositols by High Performance Liquid Chromatography
Sample and Standards Preparation
HPLC Analytical Methods
Cytosolic Free Calcium Measurements in Single Cells Using Calcium-Sensitive Fluorochromes
Membrane Permeabilization with Bacterial Toxins
Measurement of Ion Fluxes and pH Gradients Across Cell Membranes
Ligand Binding and Processing: The Perfused Liver as a Model System
The Binding of Protein-Ligands to Cell-Surface Receptors
Density Gradient Media
Balanced Salt Solutions
Index
Table of Contents provided by Publisher. All Rights Reserved.

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Biomembrane Protocols

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Biomembrane Protocols

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