JAK-STAT Signalling

Analysis of Janus Tyrosine Kinase Phosphorylation and Activation.- Co-Immunoprecipitation Protocol to Investigate Cytokine Receptor Associated Proteins e.g. Janus Kinases or Other Associated Signaling Proteins.- In Vitro JAK Kinase Activity and Inhibition Assays.- Quantitative Analysis of JAK Binding using Isothermal Titraion Calorimetry and Surface Plasmon Resonance.- Determination of Protein Turnover Rates in the JAK/STAT Pathway using a Radioactive Pulse-Chase Approach.- Designing RNAi Screens to Identify JAK/STAT Pathway Components.- The Use of JAK-Specific Inhibitors as Chemical Biology Tools.- Methods for Detecting Mutations in the Human JAK2 Gene.- Detection of Activated STAT Proteins.- Detection of Activated STAT Species Using Electrophoretic Mobility Shift Assay (EMSA) and Potential Pitfalls Arising from the Use of Detergents.- Flow Cytometric Analysis of STAT Phosphorylation.- Acetylation of Endogenous STAT Proteins.- Detection and Cellular Localization of Phospho-STAT2 in the Central Nervous System by Immunohistochemical Staining.- Nuclear Trafficking of STAT Proteins Visualized by Live Cell Imaging.- Characterization of STAT Self-Association by Analytical Ultracentrifugation.- Constitutively Active STAT5 Constructs.- Analysis of Suppressor of Cytokine Signalling (SOCS) Gene Expression by Real-Time Quantitative PCR.- Detection of Endogenous SOCS1 and SOCS3 Proteins by Immunoprecipitation and Western Blot Analysis.- In vitro Ubiquitination of Cytokine Signaling Components.- Production and Crystallization of Recombinant JAK Proteins.- Bacterial Expression, Purification and Crystallization of Tyrosine Phosphorylated STAT Proteins.