Dab Immunohistochemistry
Dab Immunohistochemistry - Related Videos
09:23
Wholemount Immunohistochemistry for Revealing Complex Brain Topography
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Cited by 17 •
2012
Neural circuits are topographically organized into functional compartments with specific molecular profiles. Here, we provide the practical and technical steps for revealing global brain topography using a versatile wholemount immunohistochemical staining approach. We demonstrate the utility of the method using the well-understood cytoarchitecture and circuitry of cerebellum.
13:24
Immunohistochemistry and Immunocytochemistry: Tissue Imaging via Light Microscopy
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2023
Source: Michael S. Lee1 and Tonya J. Webb1 1 Department of Microbiology and Immunology, University of Maryland School of Medicine and the Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, Maryland 21201 Immunohistochemistry (IHC) and immunocytochemistry (ICC) are techniques used to visualize the expression and localization of specific antigens using antibodies. The first published use of IHC was in 1941 when Albert Coons used the technique to visualize the presence of...
05:45
Quantitative Immunohistochemistry of the Cellular Microenvironment in Patient Glioblastoma Resections
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Cited by 10 •
2017
This protocol was developed to quantitatively identify tumor microenvironment components in glioblastoma patient resections using chromogenic immunohistochemistry and ImageJ.
11:27
Immunohistochemistry on Paraffin Sections of Mouse Epidermis Using Fluorescent Antibodies
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Cited by 2 •
2008
We describe a reliable method for immunolocalization within the epidermis modified for both frozen and paraffin sections that we use very routinely in our laboratory.
08:29
Drosophila Pupal Abdomen Immunohistochemistry
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Cited by 20 •
2011
Antibody staining of the Drosophila pupae can enhance genetic analyses of adult abdominal developmental genetics. We present our protocol for dissection, fixation and antibody staining of staged Drosophila pupal abdomen.
FAQs
Specificity arises from pairing the primary antibody with its target antigen in the tissue. The secondary antibody then recognizes the primary antibody and supplies the enzyme-linked detection step. This layered arrangement connects molecular recognition to a visible signal, allowing staining patterns to be interpreted in relation to particular proteins or cellular structures.
Horseradish peroxidase serves as the enzymatic link between antibody binding and DAB visualization. When hydrogen peroxide is present, the enzyme-driven reaction oxidizes DAB where the antibody complex has localized. This confines the brown reaction product to antigen-associated sites, which helps preserve spatial information for microscopic analysis of nervous system tissue.
Because oxidized DAB becomes insoluble, the reaction product remains at the antibody binding location instead of dispersing through the section. That property creates contrast against surrounding tissue and supports assessment of where a target occurs. In neuroscience sections, this spatial retention helps relate protein labeling to neuronal or glial organization and morphology.
A practical workflow begins with a tissue section containing the antigen of interest, followed by application of a primary antibody. A horseradish peroxidase-linked secondary antibody binds that primary antibody. Hydrogen peroxide then enables DAB oxidation at the labeled sites, and the resulting tissue contrast is examined microscopically to evaluate protein or cellular localization.
Microscopic evaluation can show the distribution of a labeled protein or cellular structure across a tissue section. Researchers can also examine cellular morphology and compare patterns of expression with nervous system organization. These observations support analysis of how neuronal markers, glial proteins, or disease-associated changes are arranged within brain and spinal cord tissue.
In neuroscience, the method supports mapping neuronal markers and glial proteins within brain and spinal cord sections. Researchers can use the resulting brown labeling to examine nervous system organization and identify disease-associated changes in tissue. The same spatial readout links protein expression with the distribution and morphology of cells observed under the microscope.