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.
Date Published: 1/08/2008, Issue 11; doi: 10.3791/552
Keywords: Cellular Biology, Issue 11, Springer Protocols, Immunohistochemistry, epidermis, differentiation, keratins, antibody
Troy, T., Arabzadeh, A., Enikanolaiye, A., Lariviere, N., Turksen, K. Immunohistochemistry on Paraffin Sections of Mouse Epidermis Using Fluorescent Antibodies. J. Vis. Exp. (11), e552, doi:10.3791/552 (2008).
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.
The complete text protocol for this experimental approach is available in Springer Protocols.
The authors have nothing to disclose.