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E-book

Title Hedgehog signaling protocols / edited by Natalia A. Riobo
Edition 2nd ed
Published New York : Humana Press : Springer, 2015

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Description 1 online resource (xvi, 200 pages) : illustrations (some color)
Series Methods in molecular biology ; 1322
Methods in molecular biology (Clifton, N.J.) ; v. 1322
Contents Controlled Delivery of Sonic Hedgehog with a Heparin-Based Coacervate -- In Vivo Imaging of Hedgehog Transport in Drosophila Epithelia -- Modeling Hedgehog Signaling Through Flux-Saturated Mechanism -- Evaluating the Activity of Smoothened Toward G Proteins Using [35S] Guanosine 5'-(3-O-thio)triphosphate ([35S]GTPÞS) -- Analysis of Smoothened Phosphorylation and Activation in Cultured Cells and Wing Discs of Drosophila -- Investigation of Protein-Protein Interactions and Conformational Changes in Hedgehog Signaling Pathway by FRET -- Luciferase Reporter Assays to Study Transcriptional Activity of HH Signaling in Normal and Cancer Cells -- Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR -- Quantitative Immunoblotting of Endogenous Hedgehog Pathway Components -- Measuring Gli2 Phosphorylation by Selected Reaction Monitoring Mass Spectrometry -- Rapid Screening of Gli2/3 Mutants Using the Flp-In System -- Insights into Gli Factors Ubiquitylation Methods -- Determination of Acetylation of the Gli Transcription Factors -- Efficient Detection of Indian Hedgehog During Endochondral Ossification by Whole Mount Immunofluorescence -- Methods for Detection of Ptc1-Driven LacZ Expression in Adult Mouse Skin -- Determination and Analysis of Cellular Metabolic Changes by Non-Canonical Hedgehog Signaling
Summary Annotation The second edition of this volume provides novel protocols for the study of newly discovered function and modifications of components of the Hh pathways. The first few chapters discuss methods for stable delivery of soluble Hh proteins in cell cultures that can be adapted for in vivo studies, and mathematical models for studying Hh proteins gradient formation in tissues. The next few chapters explore studies in cell culture using fibroblasts that respond readily to Hh proteins. The book goes on to present protocols for evaluation of transcriptional activity of the Gli transcription factors in cell cultures and determine the expression level of endogenous Gli proteins. The final chapters talk about metabolic effects of noncanonical Hh signaling. Written in the highly successful Methods in Molecular Biology series format, chapters include 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 authoritative, Hedgehog Signaling Protocols, Second Edition presents researchers with a comprehensive understanding of the Hh pathway and its function in development and diseases
Analysis eiwitten
proteins
Proteins and Enzymes
Eiwitten en enzymen
Bibliography Includes bibliographical references and index
Notes Online resource; title from PDF title page (SpringerProtocols, viewed July 20, 2015)
Subject Cellular signal transduction -- Research -- Methodology
Transcription factors -- Research -- Methodology
Cellular signal transduction.
Genetic regulation.
Hedgehog Proteins
Microbiological Techniques
Signal Transduction
Gene Expression Regulation
Proteins.
Science -- Life Sciences -- Biochemistry.
Genetic regulation
Cellular signal transduction
Cellular signal transduction -- Research -- Methodology
Genre/Form Laboratory manuals
Laboratory manuals.
Manuels de laboratoire.
Form Electronic book
Author Riobo, Natalia A
ISBN 9781493927722
1493927728
149392771X
9781493927715