About: Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry     Goto   Sponge   NotDistinct   Permalink

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Description
  • The TP63 gene gives rise to protein isoforms with different properties and functions due to the presence (TAp63) or absence (ΔNp63) of N-terminal p53-like transactivation domain. Immunohistochemistry for p63 has clinical value for certain tumour types, but investigations have been hampered by a lack of well characterized antibodies and the inability to discriminate between these N-terminal isoforms with opposite functional properties. We have characterized a series of monoclonal antibodies to recombinant human TAp63 and two commercial p63 monoclonals by Western blot, immunostaining and phage display epitope mapping. 28 of 29 (96.6 %) novel monoclonals that recognized all p63 isoforms showed substantial cross-reactivity with p73, as did the commercial antibody, 4A4. One novel clone, PANp63-6.1, showed slight cross-reaction with p73 by Western blotting but not immunohistochemistry and the SFI-6 monoclonal did not cross-react with p73 or p53. Phage display revealed that PANp63-6.1 epitope has one amino acid difference between p63 and p73; 4A4 epitope is identical in both, whereas SFI-6 epitope is unique to p63, accounting for these findings. We also produced and characterized TAp63-specific clone that does not recognize p53 or p73, and we prepared polyclonal sera specific for ΔNp63 isoforms. Immunohistochemistry demonstrated that TAp63 is expressed in a variety of epithelial and other cell types during development, often in a converse pattern to ΔNp63, but has a very limited expression in normal adult tissues and is independent of ΔNp63. TAp63 was expressed in 17.6 % of squamous cancers of cervix that expressed p63, unlike normal cervix where TAp63 was not expressed. TAp63 did not associate with proliferative index, but cervical carcinomas with TAp63 expression showed improved survival. These data highlight the need for rigorous antibody characterization and indicate that p63-isoform identification may improve the clinical value of p63 expression analyses.
  • The TP63 gene gives rise to protein isoforms with different properties and functions due to the presence (TAp63) or absence (ΔNp63) of N-terminal p53-like transactivation domain. Immunohistochemistry for p63 has clinical value for certain tumour types, but investigations have been hampered by a lack of well characterized antibodies and the inability to discriminate between these N-terminal isoforms with opposite functional properties. We have characterized a series of monoclonal antibodies to recombinant human TAp63 and two commercial p63 monoclonals by Western blot, immunostaining and phage display epitope mapping. 28 of 29 (96.6 %) novel monoclonals that recognized all p63 isoforms showed substantial cross-reactivity with p73, as did the commercial antibody, 4A4. One novel clone, PANp63-6.1, showed slight cross-reaction with p73 by Western blotting but not immunohistochemistry and the SFI-6 monoclonal did not cross-react with p73 or p53. Phage display revealed that PANp63-6.1 epitope has one amino acid difference between p63 and p73; 4A4 epitope is identical in both, whereas SFI-6 epitope is unique to p63, accounting for these findings. We also produced and characterized TAp63-specific clone that does not recognize p53 or p73, and we prepared polyclonal sera specific for ΔNp63 isoforms. Immunohistochemistry demonstrated that TAp63 is expressed in a variety of epithelial and other cell types during development, often in a converse pattern to ΔNp63, but has a very limited expression in normal adult tissues and is independent of ΔNp63. TAp63 was expressed in 17.6 % of squamous cancers of cervix that expressed p63, unlike normal cervix where TAp63 was not expressed. TAp63 did not associate with proliferative index, but cervical carcinomas with TAp63 expression showed improved survival. These data highlight the need for rigorous antibody characterization and indicate that p63-isoform identification may improve the clinical value of p63 expression analyses. (en)
Title
  • Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry
  • Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry (en)
skos:prefLabel
  • Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry
  • Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry (en)
skos:notation
  • RIV/00209805:_____/13:#0000423!RIV14-GA0-00209805
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(ED2.1.00/03.0101), P(GAP301/11/1678), P(NT13794)
http://linked.open...iv/cisloPeriodika
  • 3
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
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http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 65105
http://linked.open...ai/riv/idVysledku
  • RIV/00209805:_____/13:#0000423
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • p63; TAp63; development; cervical cancer (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • DE - Spolková republika Německo
http://linked.open...ontrolniKodProRIV
  • [5830AE71C65C]
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  • Virchows Archiv
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http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 463
http://linked.open...iv/tvurceVysledku
  • Mouková, Lucie
  • Vojtěšek, Bořivoj
  • Müller, Petr
  • Holčáková, Jitka
  • Nenutil, Rudolf
  • Nekulová, Miroslava
  • Bouchalová, Pavla
http://linked.open...ain/vavai/riv/wos
  • 000323904700007
issn
  • 0945-6317
number of pages
http://bibframe.org/vocab/doi
  • 10.1007/s00428-013-1459-4
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