Signal Transduction Immunohistochemistry

This book presents protocols that address the numerous challenges of signal-transduction immunohistochemistry (ST-IHC). Coverage includes protocols in digital imaging and image analysis of stained cells and techniques used in cancer and stem cell research.

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.

Presents detailed, thorough protocols examining this burgeoning field of study Features techniques easily adaptable to numerous research tasks Contains expert notes and vital implementation advice Includes supplementary material: sn.pub/extras

Inhalt

Overview of the Generation, Validation, and Application of Phosphosite-Specific Antibodies.- Selection and Validation of Antibodies for Signal Transduction Immunohistochemistry.- An Overview of Western Blotting for Determining Antibody Specificities for Immunohistochemistry.- Optimized Protocol to Make Phospho-Specific Antibodies that Work.- Methodology and Technology for Stabilization of Specific States of Signal Transduction Proteins.- An Enhanced Antigen Retrieval Protocol for Immunohistochemical Staining of Formalin-Fixed, Paraffin-Embedded Tissues.- Imaging Techniques in Signal Transduction IHC.- Practical Considerations of Image Analysis and Quantification of Signal Transduction IHC Staining.- Flow Cytometric Analysis of Cell Signaling Proteins.- CytoSys: A Tool for Extracting Cell Cycle-Related Expression Dynamics from Static Data.- Signaling Events Initiated by Kappa Opioid Receptor Activation: Quantification and Immunolocalization Using Phospho-Selective KOR, p38-MAPK, and K IR 3.1 Antibodies.- Immunohistochemical Assessment of Signal Transduction and Cell-Cycle Networks in Neural Tumors.- Novel Multi-Color Immunofluorescence Technique Using Primary Antibodies Raised in the Same Host Species.- Activation and Differentiation of Mesenchymal Stem Cells.- Double In situ Detection of Sonic Hedgehog mRNA and pMAPK Protein in Examining the Cell Proliferation Signaling Pathway in Mouse Embryo.- Identifying Intracellular Sites of Eicosanoid Lipid Mediator Synthesis with EicosaCell Assays.- Absorption Control in Immunohistochemistry Using Phospho-Peptides Immobilized on Magnetic Beads.