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Hedgehog Signaling Protocols

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Cover of 'Hedgehog Signaling Protocols'

Table of Contents

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    Book Overview
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    Chapter 1 Controlled Delivery of Sonic Hedgehog with a Heparin-Based Coacervate
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    Chapter 2 In Vivo Imaging of Hedgehog Transport in Drosophila Epithelia.
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    Chapter 3 Modeling Hedgehog Signaling Through Flux-Saturated Mechanisms
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    Chapter 4 Evaluating the Activity of Smoothened Toward G Proteins Using [ 35 S]Guanosine 5′-(3- O -thio)triphosphate ([ 35 S]GTPγS)
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    Chapter 5 Analysis of Smoothened Phosphorylation and Activation in Cultured Cells and Wing Discs of Drosophila
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    Chapter 6 Hedgehog Signaling Protocols
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    Chapter 7 Luciferase Reporter Assays to Study Transcriptional Activity of Hedgehog Signaling in Normal and Cancer Cells
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    Chapter 8 Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR
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    Chapter 9 Quantitative Immunoblotting of Endogenous Hedgehog Pathway Components
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    Chapter 10 Measuring Gli2 Phosphorylation by Selected Reaction Monitoring Mass Spectrometry
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    Chapter 11 Rapid Screening of Gli2/3 Mutants Using the Flp-In System
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    Chapter 12 Insights into Gli Factors Ubiquitylation Methods
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    Chapter 13 Determination of Acetylation of the Gli Transcription Factors
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    Chapter 14 Efficient Detection of Indian Hedgehog During Endochondral Ossification by Whole-Mount Immunofluorescence
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    Chapter 15 Methods for Detection of Ptc1-Driven LacZ Expression in Adult Mouse Skin
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    Chapter 16 Determination and Analysis of Cellular Metabolic Changes by Noncanonical Hedgehog Signaling.
Attention for Chapter 9: Quantitative Immunoblotting of Endogenous Hedgehog Pathway Components
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Chapter title
Quantitative Immunoblotting of Endogenous Hedgehog Pathway Components
Chapter number 9
Book title
Hedgehog Signaling Protocols
Published in
Methods in molecular biology, January 2015
DOI 10.1007/978-1-4939-2772-2_9
Pubmed ID
Book ISBNs
978-1-4939-2771-5, 978-1-4939-2772-2
Authors

Shohreh F. Farzan, David J. Robbins

Abstract

Quantitative analysis and modeling of signaling pathway components can reveal important information about the dynamics of that system, including the relative stoichiometries and affinities between the individual signaling components, as well as rate-limiting steps in the signaling pathway. In this chapter, we present a method that we developed to quantify the steady-state ratio of core Hedgehog (Hh) signaling components in both cultured cells and the Drosophila embryo, a physiologically relevant tissue.