Signal Transduction Immunohistochemistry: Methods and Protocols

ISBN-13: 9781617790232 / Angielski / Twarda / 2011 / 302 str.

Alexander E. Kalyuzhny

Opis

Unlike detecting constitutively expressed targets, immunohistochemical detection of labile, low abundance, and short-lived signal transduction molecules can be a very difficult task. In Signal Transduction Immunohistochemistry: Methods and Protocols, IHC experts contribute detailed protocols addressing the numerous challenges of signal-transduction immunohistochemistry (ST-IHC). Beginning with a set of introductory chapters, the volume moves on to cover techniques used for the preservation of antigens and their unmasking, protocols in digital imaging and image analysis of stained cells and tissues, high-throughput data collection and data analysis, and techniques used in neuroscience as well as cancer and stem cell research. Written in the highly successful Methods in Molecular Biology series format, chapters include brief 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. Authoritative and practical, Signal Transduction Immunohistochemistry: Methods and Protocols serves as an ideal guide for novices and as a bastion of inspiring ideas to be exploited by experienced researchers on the lookout for new experimental tricks and hints.

Recenzje

From the reviews:

"Each of us daily using immunohistochemical protocols to reveal targets either useful for research or diagnostic aims will surely wonder by which tricky techniques it is possible to overcome the preservation and unmasking of those labile antigens involved in signal transduction. Well, by seventheen chapters grouped in five parts Prof. Alexander E. Kalyuzhny is presenting an invaluable technical and methodological source of hints to satisfy our needs: to overcome troubleshottings if we are already in the field or to orientate those entering the field." (Carlo Albert Redi, European Journal of Histochemistry, July, 2011)

Spis treści

Part I: Antibodies as a Tool: From Concept to Design and Application

 

1. Overview of the Generation, Validation, and Application of Phosphosite-Specific Antibodies

            Kathy Brumbaugh, Wade Johnson, Wen-Chieh Liao, Mong-Shang Lin, J. P. Houchins, Jeff Cooper, Steven Stoesz, and Roberto Campos-Gonzalez

 

2. Selection and Validation of Antibodies for Signal Transduction Immunohistochemistry

            Juraj Bodo and Eric D. His

 

3. An Overview of Western Blotting for Determining Antibody Specificities for Immunohistochemistry

            Biji T. Kurien, Yaser Dorri, Skyler Dillon, Anil Dsouza, and R. Hal Scofield

 

4. Optimized Protocol to Make Phospho-Specific Antibodies that Work

            Amy J. Archuleta, Crystal A. Stutzke, Kristin M. Nixon, and Michael D. Browning

 

Part II: Preservation and Unmasking of Tissue Antigens

 

5. Methodology and Technology for Stabilization of Specific States of Signal Transduction Proteins

            Mats Borén

 

6. An Enhanced Antigen Retrieval Protocol for Immunohistochemical Staining of Formalin-Fixed, Paraffin-Embedded Tissues

            Sergei I. Syrbu and Michael B. Cohen

 

Part III: Imaging Techniques and High-Throughput Data Analysis

 

7. Imaging Techniques in Signal Transduction IHC

            Jerry Sedgewick

 

8. Practical Considerations of Image Analysis and Quantification of Signal Transduction IHC Staining

            Michael Grunkin, Jakob Raundahl, and Niels T. Foged

 

9. Flow Cytometric Analysis of Cell Signaling Proteins

            Maria A. Suni and Vernon C. Maino

 

10. CytoSys: A Tool for Extracting Cell Cycle-Related Expression Dynamics from Static Data

            Jayant Avva, Michael C. Weis, Radina P. Soebiyanto, James W. Jacobberger, and Sree N. Sreenath

 

Part IV: Neuroscience, Cancer, and Stem Cell Research

 

11. Signaling Events Initiated by Kappa Opioid Receptor Activation: Quantification and Immunolocalization Using Phospho-Selective KOR, p38-MAPK, and K_IR 3.1 Antibodies

            Julia C. Lemos, Clarisse Roth, and Charles Chavkin

 

12. Immunohistochemical Assessment of Signal Transduction and Cell-Cycle Networks in Neural Tumors

            Daniel Ciznadija, Afsar Barlas, and Katia Manova

 

13. Novel Multi-Color Immunofluorescence Technique Using Primary Antibodies Raised in the Same Host Species

            Jillian Frisch, J.P. Houchins, Michael Grahek, Jordan Schoephoerster, Jodi Hagen, Joseph Sweet, Leopoldo Mendoza, David Schwartz, and Alexander E. Kalyuzhny

 

14. Activation and Differentiation of Mesenchymal Stem Cells

            Pravin J. Mishra and Debabrata Banerjee

 

Part V: Novel Assays and Techniques

 

15. Double In situ Detection of Sonic Hedgehog mRNA and pMAPK Protein in Examining the Cell Proliferation Signaling Pathway in Mouse Embryo

            Sho Fujisawa, Mesruh Turkekul, Afsar Barlas, Ning Fan, and Katia Manova

 

16. Identifying Intracellular Sites of Eicosanoid Lipid Mediator Synthesis with EicosaCell Assays

            Christianne Bandeira-Melo¹, Peter F. Weller², and Patricia T. BozzaChristianne Bandeira-Melo, Peter F. Weller, and Patricia T. Bozza

 

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

            Jordan Schoephoerster, Jillian Frisch, Michael Grahek, Chun Wu, Yingwei He, Wei Wang, Jennifer Nguyen, David Schwartz, and Alexander E. Kalyuzhny