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Signal Transduction Immunohistochemistry

Methods and Protocols

  • Book
  • © 2017

Overview

Editors:
  1. Alexander E. Kalyuzhny

    1. Immunocytochemistry & Elispot Assays, Bio-Techne, Inc., Minneapolis, USA

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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 1554)

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Table of contents (21 protocols)

  1. Front Matter

    Pages i-x
    Download chapter PDF

  2. Phosphosite-Specific Antibodies: A Brief Update on Generation and Applications

    • Kathy Brumbaugh, Wen-Chie Liao, J. P. Houchins, Jeff Cooper, Steve Stoesz
    Pages 1-40

  3. Antibodies That Work Again and Again and Again

    • Kameron Simpson, Michael Browning
    Pages 41-59

  4. Validating Antibody Specificities for Immunohistochemistry by Protein Blotting Methods

    • Biji T. Kurien, R. Hal Scofield
    Pages 61-73

  5. Acquisition and Post-Processing of Immunohistochemical Images

    • Jerry Sedgewick
    Pages 75-106

  6. Using Digital Quantification of Stained Tissue Microarrays as a Medium-Throughput, Quantitative Method for Measuring the Kinetics of Signal Transduction

    • Jessica Kalra, Jennifer Baker
    Pages 107-125

  7. EicosaCell: An Imaging-Based Assay to Identify Spatiotemporal Eicosanoid Synthesis

    • Christianne Bandeira-Melo, Ligia Almeida Paiva, Natália R. T. Amorim, Peter F. Weller, Patricia T. Bozza
    Pages 127-141

  8. Preservation of Specific Protein Signaling States Using Heat Based Stabilizor System

    • Mats Borén
    Pages 143-153

  9. Hapten–Anti-Hapten Technique for Two-Color IHC Detection of Phosphorylated EGFR and H2AX Using Primary Antibodies Raised in the Same Host Species

    • Jodi Hagen, David Schwartz, Alexander E. Kalyuzhny
    Pages 155-160

  10. Culture, Transfection, and Immunocytochemical Analysis of Primary Macrophages

    • Charles A. Warwick, Yuriy M. Usachev
    Pages 161-173

  11. Phenotyping CD4+ hTh2 Cells by Flow Cytometry: Simultaneous Detection of Transcription Factors, Secreted Cytokines, and Surface Markers

    • Chris Goetz, Li-Jen Peng, Birte Aggeler, Jody Bonnevier
    Pages 175-184

  12. A Coupled Approach Utilizing Immunohistochemistry and Immunocytochemistry to Visualize Cellular Lipophagy

    • Aishwarya Sathyanarayan
    Pages 185-191

  13. Apoptotic Bodies: Selective Detection in Extracellular Vesicles

    • Paul Hauser, Sha Wang, Vladimir V. Didenko
    Pages 193-200

  14. Activation and Differentiation of Mesenchymal Stem Cells

    • Pravin J. Mishra, Debabrata Banerjee
    Pages 201-209

  15. High-Sensitivity IHC Detection of Phosphorylated p27/Kip1 in Human Tissues Using Secondary Antibody Conjugated to Polymer-HRP

    • Michael Grahek, Ana Ptak, Alexander E. Kalyuzhny
    Pages 211-218

  16. Using Phospho-Peptides Immobilized on Magnetic Beads for Absorption Control in Immunohistochemistry

    • David Schwartz, Michael Grahek, Yingwei He, Wei Wang, Jennifer Nguyen, Alexander E. Kalyuzhny
    Pages 219-227

  17. Dual Detection of Nucleolytic and Proteolytic Markers of Lysosomal Cell Death: DNase II-Type Breaks and Cathepsin D

    • Candace L. Minchew, Vladimir V. Didenko
    Pages 229-236

  18. Multiplex Immunohistochemistry for Mapping the Tumor Microenvironment

    • Jessica Kalra, Jennifer Baker
    Pages 237-251

  19. Validation of Anti-Mouse PDL-1 Goat Polyclonal Antibody Staining with Mouse PDL-1 In Situ Hybridization on Adjacent Sections of Cell Pellets and Mouse Tumors

    • Jacob Ricca, Mesruh Turkekul, Afsar Barlas, Dmitry Yarilin, Sho Fujisawa, Ning Fan et al.
    Pages 253-262

  20. Automated Double In Situ Detection of Mouse Lgr5 mRNA and Lysozyme Protein in Examining the Neighboring Cell Types of the Mouse Intestinal Crypt

    • Mesruh Turkekul, Afsar Barlas, Dmitry Yarilin, Sho Fujisawa, Ning Fan, Matthew Brendel et al.
    Pages 263-272
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Keywords

About this book

This second edition volume provides detailed protocols that address the challenges of signal-transduction IHC. This book delves into chapters that discuss the nature of signal transduction phenomena and approaches to making phosphor-specific antibodies, as well as numerous bona fide methods methods on digital imaging techniques, preservation of tissue targets, multicolor detection, flow cytometry, lipophagy analysis, apoptosis, and the combination of IHC with in situ hybridization. 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.

Thorough and comprehensive, Signal Transduction Immunohistochemistry: Methods and Protocols, Second Edition is a valuable resource to both novices and experts in other fields of biomedical research who need advice on IHC protocols to study signal transduction. This book will also be useful for researchers in academia, government labs, and the biotech industry.

Editors and Affiliations

  • Immunocytochemistry & Elispot Assays, Bio-Techne, Inc., Minneapolis, USA

    Alexander E. Kalyuzhny

Bibliographic Information

  • Book Title: Signal Transduction Immunohistochemistry

  • Book Subtitle: Methods and Protocols

  • Editors: Alexander E. Kalyuzhny

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-6759-9

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media LLC 2017

  • Hardcover ISBN: 978-1-4939-6757-5Published: 21 February 2017

  • Softcover ISBN: 978-1-4939-8294-3Published: 20 June 2018

  • eBook ISBN: 978-1-4939-6759-9Published: 07 February 2017

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 2

  • Number of Pages: X, 290

  • Number of Illustrations: 22 b/w illustrations, 45 illustrations in colour

  • Topics: Immunology

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