Immunofluorescence microscopy for imaging of nuclear p63 in human primary keratinocytes: a comparison of antibodies and fixation methods.
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.
Markert
Chad D
CD
Wake Forest Institute for Regenerative Medicine, United States. cmarkert@wfubmc.edu
Bharadwaj
Shantaram
S
Zhang
Yuanyuan
Y
Childers
Martin K
MK
Furth
Mark E
ME
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
D000906:Antibodies; C081876:CKAP4 protein, human; D008565:Membrane Proteins; D009687:Nuclear Proteins