Signal transduction immunohistochemistry : methods and protocols

Author(s)

    • Kalyuzhny, Alexander E.

Bibliographic Information

Signal transduction immunohistochemistry : methods and protocols

edited by Alexander E. Kalyuzhny

(Methods in molecular biology / John M. Walker, series editor, 1554)(Springer protocols)

Humana Press, c2017

2nd ed

Available at  / 1 libraries

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Note

Includes bibliographical references and index

Description and Table of Contents

Description

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.

Table of Contents

Preface... Table of Contents... Contributing Authors... 1. Phosphosite-Specific Antibodies: A Brief Update on Generation and Applications Kathy Brumbaugh, Wen-Chie Liao, J. P. Houchins, Jeff Cooper, and Steve Stoesz 2. Antibodies That Work Again and Again and Again Kameron Simpson and Michael Browning 3. Validating Antibody Specificities for Immunohistochemistry by Protein Blotting Methods Biji T. Kurien and R. Hal Scofield 4. Acquisition and Post-Processing of Immunohistochemical Images Jerry Sedgewick 5. Using Digital Quantification of Stained Tissue Microarrays as a Medium-Throughput, Quantitative Method for Measuring the Kinetics of Signal Transduction Jessica Kalra and Jennifer Baker 6. EicosaCell: An Imaging-Based Assay to Identify Spatiotemporal Eicosanoid Synthesis Christianne Bandeira-Melo, Ligia Almeida Paiva, Natalia R. T. Amorim, Peter F. Weller, and Patricia T. Bozza 7. Preservation of Specific Protein Signaling States using Heat Based Stabilizor System Mats Boren 8. 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, and Alexander E. Kalyuzhny 9. Culture, Transfection and Immunocytochemical Analysis of Primary Macrophages Charles A. Warwick and Yuriy M. Usachev 10. Phenotyping CD4+ hTh2 Cells by Flow Cytometry: Simultaneous Detection of Transcription Factors, Secreted Cytokines and Surface Markers Chris Goetz, Li-Jen Peng, Birte Aggeler, and Jody Bonnevier 11. A Coupled Approach Utilizing Immunohistochemistry and Immunocytochemistry to Visualize Cellular Lipophagy Aishwarya Sathyanarayan 12. Apoptotic Bodies: Selective Detection in Extracellular Vesicles Paul Hauser, Sha Wang, and Vladimir V. Didenko 13. Activation and Differentiation of Mesenchymal Stem Cells Pravin J. Mishra and Debabrata Banerjee 14. High-Sensitivity IHC Detection of Phosphorylated p27/Kip1 in Human Tissues using Secondary Antibody Conjugated to Polymer-HRP Michael Grahek, Ana Ptak, and Alexander E. Kalyuzhny 15. Using Phospho-Peptides Immobilized on Magnetic Beads for Absorption Control in Immunohistochemistry David Schwartz, Michael Grahek, Yingwei He, Wei Wang, Jennifer Nguyen, and Alexander E. Kalyuzhny 16. Dual Detection of Nucleolytic and Proteolytic Markers of Lysosomal Cell Death: DNase II-Type Breaks and Cathepsin D Candace C. Minchew and Vladimir V. Didenko 17. Multiplex Immunohistochemistry for Mapping the Tumor Microenvironment Jessica Kalra and Jennifer Baker 18. 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, Matthew Brendel, Dmitriy Zamarin, Jedd D. Wolchok, and Katia Manova-Todorova 19. 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, and Katia Manova-Todorova 20. Automated High Content Screening of g--H2AX Expression in HeLa Cells Nate Hopp, Jodi Hagen, Birte Aggeler, and Alexander E. Kalyuzhny

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