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Biomembrane Protocols, Volume 2: Architecture and Function

Handbook / Manual

Series: Methods in Molecular Biology Volume: 27

Edited By: JM Graham and JA Higgins

380 pages, 55 illus

Humana Press

Spiralbound | Dec 1994 | #33060 | ISBN: 0896032507
Availability: Usually dispatched within 1-2 weeks Details
NHBS Price: £64.00 $81/€76 approx

About this book

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.

...the editors have assembled an authoritative set of reviews, which give generally clear practical guidance and useful methodological tips. - SGM Quarterly


Contents

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: Part A: Labeling of Oxidized Glycoproteins with 3H-Borohydride; Part B: Identification and Separation of Integral Membrane Proteins Using Triton X-114; Part C: 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: Part A: Preparation of Antibodies; Part B: Affinity-Purification of Antipeptide Antibodies; Part C: Purification of Membrane Proteins by Immunoaffinity Chromatography; Part D: Competitive ELISA for Determining Membrane Protein Topology. The Production of Monoclonal Antibodies to Membrane Proteins: Part A: Production of Hybridoma Cell Lines; Part B: 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: Part A: Synthesis of Fluorescent Glycerolipid Probes; Part B: 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: Part A: Identification of G-Proteins; Part B: GTPase Studies; Part C: Reconstitution of Second Messenger Pathways in Permeabilized Cell Preparations Using GTP Analogs and Receptor Ligands. Assay of Protein Kinases and Protein Phosphorylation: Part A: Protein Kinase-Mediated Phosphorylation Events; Part B: Measurement of Protein Kinase Expression; Part C: Measurement of Protein Kinase Activity and Translocation. Analysis of Cellular Phosphoinositides and Phosphoinositols: Extraction and Simple Analytical Procedures: Part A: Biosynthesis and Extraction of Phosphoinositides and Phosphoinositols; Part B: Separation of Phosphoinositols by Anion-Exchange Chromatography; Part C: Separation of Phosphoinositides by Thin-Layer Chromatography; Part D: Deacylation of Phospholipids and Separation of Products by Anion-Exchange Chromatography. Analysis of Cellular Phosphoinositides and Phosphoinositols by High Performance Liquid Chromatography: Part A: Sample and Standards Preparation; Part B: 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. Appendix: 1. Density Gradient Media. Appendix: 2. Balanced Salt Solutions. Index.

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