This detailed book covers numerous expert methodologies to examine Hippo signaling on the structural, molecular, cellular, and organismal level. Beginning with a section on Drosophila genetics, the volume continues with parts exploring molecular and cell biological studies of the Hippo pathway, the use of structural biology and biochemistry, and a brief look at the Hippo pathway in mouse models. Written for the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily…mehr
This detailed book covers numerous expert methodologies to examine Hippo signaling on the structural, molecular, cellular, and organismal level. Beginning with a section on Drosophila genetics, the volume continues with parts exploring molecular and cell biological studies of the Hippo pathway, the use of structural biology and biochemistry, and a brief look at the Hippo pathway in mouse models. Written for the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, The Hippo Pathway: Methods and Protocols equips newcomers and specialists alike with key methodologies to accurately define the status of Hippo signaling in their experimental settings.
The Power of Drosophila Genetics: The Discovery of the Hippo Pathway.- Drosophila Genetics: The Power of Genetic Mosaic Approaches.- DrosophilaGenetics: Analysis of Tissue Growth in Adult Tissues.- Live Imaging of Hippo Pathway Components in Drosophila Imaginal Discs.- Localization of Hippo Signaling Components in Drosophila by Fluorescence and Immunofluorescence.- Bimolecular Fluorescence Complementation (BiFC) in Tissue Culture and in Developing Tissues of Drosophilato Study Protein-Protein Interactions.- Immunohistochemistry to Study YAP in Human Tissue Samples.-Immunofluorescence Study of Endogenous YAP in Mammalian Cells.- Immunofluorescence Microscopy to Study Endogenous TAZ in Mammalian Cells.- Nuclear/Cytoplasmic Fractionation to Study Hippo Effectors.- Luciferase Reporter Assays to Determine YAP/TAZ Activity in Mammalian Cells.- Quantitative Real-Time PCR to Measure YAP/TAZ Activity in Human Cells.- HTRF® Total and Phospho-YAP(Ser127) Cellular Assays.- Studying YAP-Mediated 3D Morphogenesis Using Fish Embryos and Human Spheroids.- Regulation of YAP/TAZ Activity by Mechanical Cues: An Experimental Overview.- CRISPR-Mediated Approaches to Regulate YAP/TAZ Levels.- Hippo Pathway Regulation by Tyrosine Kinases.- Structural and Biochemical Analyses of the Core Components of the Hippo Pathway.- Isothermal Titration Calorimetry Assays to Measure Binding Affinities In Vitro.- GST Pull-Down Assay to Measure Complex Formations.- Determining the Phosphorylation Status of Hippo Components YAP and TAZ Using Phos-tag.- Quantifying the Kinase Activities of MST1/2.- Measuring the Kinase Activities of the LATS/NDR Protein Kinases.- MST1/2 Kinase Assays Using Recombinant Proteins.- Visualizing HIPPO Signaling Components in Mouse Early Embryonic Development.- The Hippo Signaling Pathway in Regenerative Medicine.
The Power of Drosophila Genetics: The Discovery of the Hippo Pathway.- Drosophila Genetics: The Power of Genetic Mosaic Approaches.- DrosophilaGenetics: Analysis of Tissue Growth in Adult Tissues.- Live Imaging of Hippo Pathway Components in Drosophila Imaginal Discs.- Localization of Hippo Signaling Components in Drosophila by Fluorescence and Immunofluorescence.- Bimolecular Fluorescence Complementation (BiFC) in Tissue Culture and in Developing Tissues of Drosophilato Study Protein-Protein Interactions.- Immunohistochemistry to Study YAP in Human Tissue Samples.-Immunofluorescence Study of Endogenous YAP in Mammalian Cells.- Immunofluorescence Microscopy to Study Endogenous TAZ in Mammalian Cells.- Nuclear/Cytoplasmic Fractionation to Study Hippo Effectors.- Luciferase Reporter Assays to Determine YAP/TAZ Activity in Mammalian Cells.- Quantitative Real-Time PCR to Measure YAP/TAZ Activity in Human Cells.- HTRF® Total and Phospho-YAP(Ser127) Cellular Assays.- Studying YAP-Mediated 3D Morphogenesis Using Fish Embryos and Human Spheroids.- Regulation of YAP/TAZ Activity by Mechanical Cues: An Experimental Overview.- CRISPR-Mediated Approaches to Regulate YAP/TAZ Levels.- Hippo Pathway Regulation by Tyrosine Kinases.- Structural and Biochemical Analyses of the Core Components of the Hippo Pathway.- Isothermal Titration Calorimetry Assays to Measure Binding Affinities In Vitro.- GST Pull-Down Assay to Measure Complex Formations.- Determining the Phosphorylation Status of Hippo Components YAP and TAZ Using Phos-tag.- Quantifying the Kinase Activities of MST1/2.- Measuring the Kinase Activities of the LATS/NDR Protein Kinases.- MST1/2 Kinase Assays Using Recombinant Proteins.- Visualizing HIPPO Signaling Components in Mouse Early Embryonic Development.- The Hippo Signaling Pathway in Regenerative Medicine.
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