Protein Kinase Protocols

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.