Immunohistochemistry on Paraffin Sections of Mouse Epidermis Using Fluorescent Antibodies
Tammy-Claire Troy1, Azadeh Arabzadeh2, Adebola Enikanolaiye2, Nathalie Lariviere2, Kursad Turksen2 1Chronic Disease Program, Ottawa Health Research Institute, 2Department of Cellular and Molecular Medicine, University of Ottawa
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0:00 Title0 0:32 Introduction32 1:34 Mouse Dissection94 3:16 Fixation and Paraffin Block Preparation196 5:16 Dewaxing Paraffin Sections and Antigen Retrieval316 7:28 Immunohistochemistry448 10:33 Visualization of Stained Slides633 11:01 Conclusion661
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In the epidermis, immunohistochemistry is an efficient means of localizing specific proteins to their relative expression compartment; namely the basal, suprabasal, and stratum corneum layers. The precise localization within the epidermis of a particular protein lends clues toward its functional role within the epidermis. In this chapter, 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. Paraffin sections generally provide much better morphology, hence, superior results and photographs; however, not all antibodies will work with the harsh fixation and treatment involved in their processing. Therefore, the protocol for frozen sectioning is also included. Within paraffin sectioning, two fixation protocols are described (Bouin's and paraformaldehyde); the choice of fixative will be directly related to the antibody specifications and may require another fixing method.
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The complete text protocol for this experimental approach is available in Springer Protocols.
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Troy TC, Arabzadeh A, Enikanolaiye A, Lariviere N, Turksen K (2008). Immunohistochemistry on Paraffin Sections of Mouse Epidermis Using Fluorescent Antibodies. JoVE. 11. http://www.jove.com/index/details.stp?id=552, doi: 10.3791/552
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Allowed tags: i, b, u, sup, sub 
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| | 07/06/2008 12:20:45 PM
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afiat_b responded with a statement of type: Question
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| | 11/14/2009 10:44:42 AM
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mohammad ali responded with a statement of type: Question
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Mailer.URL: http://www.jove.com/index/details.stp?id=552 |
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