title: Immunofluorescence microscopy for imaging of nuclear p63 in human primary keratinocytes: a comparison of antibodies and fixation methods.
abstract: The ability of an experimental treatment to induce primitive, undifferentiated stem cells towards an epidermal fate may be tested by comparing the treated stem cells with a positive control, such as primary keratinocytes. In an effort to perfect methods used for this comparison, we tested two commercially available antibodies and three fixation methods to determine which antibody/fixation interaction produced the best immunofluorescent images of the nuclear localization of p63, a canonical marker of epidermal fate, in keratinocytes. Here, we report the methods used, and the experimental outcome.
authors.affiliation: Wake Forest Institute for Regenerative Medicine, United States. cmarkert@wfubmc.edu
authors.forename: Chad D
authors.initials: CD
authors.lastname: Markert
authors.forename: Shantaram
authors.initials: S
authors.lastname: Bharadwaj
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Martin K
authors.initials: MK
authors.lastname: Childers
authors.forename: Mark E
authors.initials: ME
authors.lastname: Furth
mesh_terms: D000906:Antibodies / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D052816:Foreskin / Q000473:pathology; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000276:immunology / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009687:Nuclear Proteins / Q000096:biosynthesis*; D039904:Pluripotent Stem Cells / Q000276:immunology / Q000378:metabolism*; D012015:Reference Standards; D016707:Tissue Fixation
chemical_list: D000906:Antibodies; C081876:CKAP4 protein, human; D008565:Membrane Proteins; D009687:Nuclear Proteins