Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry

摘要: The TP63 gene gives rise to protein isoforms with different properties and functions due the presence (TAp63) or absence (ΔNp63) of an N-terminal p53-like transactivation domain. Immunohistochemistry for p63 has clinical value certain tumour types, but investigations have been hampered by a lack well characterized antibodies inability discriminate between these opposite functional properties. We extensively series monoclonal recombinant human TAp63 two commercial monoclonals Western blot, immunostaining phage display epitope mapping. Twenty-eight 29 (96.6 %) novel that recognized all showed substantial cross-reactivity p73, as did antibody, 4A4. One clone, PANp63-6.1, slight cross-reaction p73 blotting not immunohistochemistry SFI-6 cross-react p53. Phage revealed PANp63-6.1 one amino acid difference 4A4 is identical in both, whereas unique p63, accounting findings. also produced TAp63-specific clone does recognize p53 we prepared polyclonal sera specific ΔNp63 isoforms. demonstrated expressed variety epithelial other cell types during development, often converse pattern ΔNp63, very limited expression normal adult tissues independent ΔNp63. was 17.6 % squamous cancers cervix unlike where expressed. associate proliferative index, cervical carcinomas improved survival. These data highlight need rigorous antibody characterization indicate p63-isoform identification may improve analyses.