Overview

In Protein Kinase Protocols, a panel of highly skilled laboratory investigators describe both basic and more sophisticated methods for the analysis of kinase-mediated signaling cascades, with emphasis on the identification of proteins according to their interactive relationships and the analysis of their functional properties. Described in step-by-step detail, these readily reproducible techniques offer novices quick access to a complicated field, and more experienced investigators many novel and time-saving ploys. Emphasis is given to critical technical steps that are often omitted from methods published in the primary literature. Broad in its range of techniques and thoroughly detailed to help ensure experimental success, Protein Kinase Protocols offers both novice and experienced investigators powerful tools for understanding the functional roles of specific protein kinases within signaling cascades and for identification and evaluation of novel therapeutic targets.

Full Product Details

Author:   Alastair D. Reith
Publisher:   Humana Press Inc.
Imprint:   Humana Press Inc.
Edition:   2001 ed.
Volume:   124
Dimensions:   Width: 15.20cm , Height: 2.50cm , Length: 22.90cm
Weight:   0.728kg
ISBN:  

9780896037007

ISBN 10:   0896037002
Pages:   365
Publication Date:   28 September 2000
Audience:   College/higher education ,  Professional and scholarly ,  Undergraduate ,  Professional & Vocational
Format:   Hardback
Publisher's Status:   Active
Availability:   In Print  
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Table of Contents

Protein Kinase-Mediated Signaling Networks.- Cloning Protein Tyrosine Kinases by Screening cDNA Libraries with Antiphosphotyrosine Antibodies.- Immunoprecipitation and Western Blotting of Phosphotyrosine-Containing Proteins.- Two-Dimensional Phosphoamino Acid Analysis.- Two-Dimensional Phosphopeptide Mapping of Receptor Tyrosine Kinases.- Identification of the Sites of Phosphorylation in Proteins Using High Performance Liquid Chromatography and Mass Spectrometry.- Phosphorylation of Smad Signaling Proteins by Receptor Serine/Threonine Kinases.- Assays for Mitogen-Activated Protein Kinase (MARK) Subtypes and MARK Activating Protein Kinase-2 (MAPKAP K-2) Using a Common Cell Lysate.- JAK-Mediated Phosphorylation and Activation of STAT Signaling Proteins.- Assays for Glycogen Synthase Kinase-3(GSK-3).- Cyclin-Dependent Kinases and Cyclin-Dependent Kinase Inhibitors.- Protein Histidine Kinase.- Cloning and Expression of Recombinant HEXA-HIS Tagged ZAP-70 Using the Baculovirus System.- Analysis of Protein Kinase Subcellular Localization by Visualization of GFP Fusion Proteins.- Detection of Phosphorylation-Dependent Interactions by Far-Western Gel Overlay.- Purification of Tyrosine-Phosphorylated Proteins by Immunoaffinity Chromatography and Direct Cloning of Their cDNAs from Bacterial Expression Libraries.- Identification of Receptor Tyrosine Kinase (RTK) Substrates by the Cloning of ReceptorTargets (CORT) Strategy.- Identification of Receptor Tyrosine Kinase Associating Proteins Using the Yeast Two-Hybrid System.- cDNA Expression Cloning and Characterization of Phosphorylation Dependent Protein Interactors Using the Yeast Tribrid System.- Analysis of SH2 Domain - Phosphopeptide Interactions by Isothermal Titration Calorimetry and Surface Plasmon Resonance.- Cloning and Characterization of RTK Ligands Using Receptor-Alkaline Phosphatase Fusion Proteins.- Isolation and Characterization of “Orphan-RTK” Ligands Using an Integrated Biosensor Approach.

Reviews

By engaging high-ranking experts as authors of the individual chapters the editor has succeeded in compiling an updated an highly useful compendium of today's technology in protein kinase research. besides its topicality, this book has the particular advantage that the chapters are arranged according to a constantly recurring scheme, that is, beginning with a comprehensive introduction, which provides the appropriate key references, and then continuing with very detailed experimental instructions, which are given clearly enough to enable even a less experienced experimentator to follow. The Notes at the end of each chapter are extremely helpful, in which the experimental steps are critically examined and explained in detail....for students and newcomers in the field it provides an excellent introduction to the methodology of protein kinase research, and for the experienced scientist it will be a highly valuable work of reference and an indispensible companion in daily laboratory work. - Chembiochem

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