Expression and analysis of exogenous proteins in epidermal cells.
In this chapter we review protocols for transient transfection of primary keratinocytes. The ability to transfect primary epidermal cells regardless of their differentiation status allows the biochemical and molecular characterization of multiple proteins. We review methods to analyze exogenous protein abundance in transfected keratinocytes by immunoblot and immunoprecipitation. We also present protocols to determine the subcellular distribution of these proteins by indirect immunofluorescence microscopy approaches.
Dagnino
Lina
L
Department of Physiology & Pharmacology, Child Health Research Institute and Lawson Health Research Institute, University of Western Ontario, London, ON, Canada.
Ho
Ernest
E
Chang
Wing Y
WY
D018797:Cell Cycle Proteins / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D015151:Immunoblotting; D047468:Immunoprecipitation; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D014162:Transfection
D018797:Cell Cycle Proteins; D011506:Proteins