Protein Kinase Protocols

Cover
Alastair D. Reith
Springer Science & Business Media, 02.02.2008 - 365 Seiten
As key components of many cell signaling pathways, protein kinases are implicated in a broad variety of diseases, including cancers and neurodegenerative conditions, and offer considerable potential as tractable targets for therapeutic intervention. 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 provide more experienced investigators many novel time-saving ploys. Emphasis is given to the critical technical steps that are often omitted from methods published in the primary literature. There are also tips on potential pitfalls and copious notes on how to adjust the protocols to work in related systems.
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.

 

Inhalt

Regulation
1
Cloning Protein Tyrosine Kinases by Screening cDNA Libraries
21
Immunoprecipitation and Western Blotting of Phosphotyrosine
39
Peter BlumeJensen and Tony Hunter
49
TwoDimensional Phosphopeptide Mapping of Receptor
67
Identification of the Sites of Phosphorylation in Proteins Using High
87
Phosphorylation of Smad Signaling Proteins by Receptor Serine
107
a Common Cell Lysate
121
Analysis of Protein Kinase Subcellular Localization
195
by FarWestern Gel Overlay
209
Purification of TyrosinePhosphorylated Proteins
221
by the Cloning of Receptor Targets CORT Strategy
239
Identification of Receptor Tyrosine Kinase Associating Proteins
251
cDNA Expression Cloning and Charaterization of Phosphorylation
271
Analysis of SH2 DomainPhosphopeptide Interactions
295
Cloning and Characterization of RTK Ligands Using Receptor
313

JAKMediated Phosphorylation and Activation of STAT Signaling
131
Darren Cross
147
Harry R Matthews and Karina Chan
171
Cloning and Expression of Recombinant HEXAHIS Tagged
183
Isolation and Characterization of OrphanRTK Ligands Using
335
Index
359
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