Signal Transduction Immunohistochemistry Methods and Protocols

This new edition collects diverse protocols compiled by experts from different areas of research as well as by biotech researchers developing novel technologies in the area of immunohistochemistry (IHC). After a few chapters to help orient novices entering the biomedical arena, the book includes methods chapters covering multiplex IHC including fluorescence and chromogenic techniques, combination of IHC with In Situ Hybridization (ISH), wavelet transform approach for organelle tracking, transcription factors in human stem cells, differentiation of mesenchymal stem cells, 3D imaging, phenotyping of glial cells in the human brain, tissue fixation, permeabilization, and cryo-preservation, as well as other topics. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Top 5 Considerations When Choosing an Antibody.- Validating Antibody Specificities for Immunohistochemistry by Protein Blotting.- Recent Advances in Chromogens for Immunohistochemistry.- Overview of Imaging Techniques in Immunohistochemistry.- Visualizing Activated Myofibroblasts Resulting from Differentiation of Mesenchymal Stem Cells.- Clinical Applications of Flow Cytometry in Cancer Immunotherapies: From Diagnosis to Treatments.- Labeling of Phospho-Specific Antibodies with oYo-Link® Epitope Tags for Multiplex Immunostaining.- Multiplex Immunochromogenic Tissue Staining Employing Primary Antibodies from the Same Species.- 3D Imaging for Cleared Tissues and Thicker Samples on Confocal and Light-Sheet Microscopes.- Optical Tissue Clearing Enables Three-Dimensional Morphometry in Experimental Nerve Regeneration Research.- Protocol and Software for Automated Detection of Lysosome Active “Runs” and “Flights” with Wavelet Transform Approach.- Phenotyping of M1 and M2a Macrophages and Differential Expression of ACE-2 on Monocytes by Flow Cytometry: Impact of Cell Culture Conditions and Sample Processing.- In Situ Hybridization (ISH) Combined with Immunocytochemistry (ICC) Co-Detection of Phosphorylated EGFR in A431 Cultured Cells.- In Situ Hybridization (ISH) Combined with Immunohistochemistry (IHC) for Co-Detection of EGFR RNA and Phosphorylated EGFR Protein in Lung Cancer Tissue.- Multiplex Spatial Protein Detection by Combining Immunofluorescence with Immunohistochemistry.- Transcription Factor-Mediated Differentiation of Motor Neurons from Human Pluripotent Stem Cells.- Immunohistochemical Techniques for Phosphoproteins.- Visualizing Cofilin-Actin Filaments by Immunofluorescence and CryoEM: Essential Steps for Observing Cofilactin in Cells.- Cyclic Multiplex Fluorescent Immunohistochemistry Protocol to Phenotype Glial Cells in Human Formalin-Fixed Paraffin-Embedded Brain Sections.- Spatially Characterizing the Immune Contexture in Mouse Tissue Using Multiplex Immunohistochemistry.- Highly Multiplexed Immuno-Fluorescence Imaging for Quantitative Spatial Analysis in Tissue Samples with ChipCytometry^TM.