title: A comprehensive test system to identify suitable antibodies against p53 for immunohistochemical analysis of canine tissues.
abstract: The tumour suppressor p53 is commonly detected in tissues of companion animals by means of antibodies raised against the human protein. The following three-step procedure was devised to test the suitability of such antibodies for immunohistochemistry on canine tissues. (1) Western blot and immunohistochemical analyses on bacterially expressed recombinant canine protein to assess human-to-canine cross-reactivity. (2) Immunohistochemistry of cultured, UVB-irradiated canine keratinocytes to evaluate suitability for detection of endogenous p53. (3) Immunohistochemistry on tissue arrays to further substantiate suitability of the antibodies on a panel of normal and neoplastic human and canine tissues. Five of six antibodies cross-reacted with recombinant canine p53. Three of these (PAb122, PAb240, CM-1) also immunolabelled stabilized wild type p53 in cell cultures and elicited a consistent, characteristic labelling pattern in a subset of tumours. However, two alternative batches of polyclonal antibody CM-1 failed to detect p53 in cell cultures, while showing a characteristic labelling pattern of a completely different subset of tumours and unspecific labelling of normal tissues. The test system described is well suited to the selection of antibodies for immunohistochemical p53 detection. The results emphasize the need to include appropriate controls, especially for polyclonal antibodies.
authors.affiliation: Institute of Veterinary Pathology.
authors.forename: S M
authors.initials: SM
authors.lastname: Keller
authors.affiliation: Institute of Veterinary Pathology.
authors.forename: B
authors.initials: B
authors.lastname: Schade
authors.affiliation: Institute of Veterinary Pathology.
authors.forename: A B
authors.initials: AB
authors.lastname: Rickenbacher
authors.affiliation: Institute of Veterinary Pathology.
authors.forename: E
authors.initials: E
authors.lastname: Brugnera
authors.affiliation: Section of Diagnostic Imaging and Radio-oncology, Vetsuisse Faculty of Zurich, Zurich.
authors.forename: M C
authors.initials: MC
authors.lastname: Wergin
authors.affiliation: Molecular Dermatology, Institute for Animal Pathology, Vetsuisse Faculty of Berne, Berne, Switzerland.
authors.forename: E J
authors.initials: EJ
authors.lastname: Müller
authors.affiliation: Molecular Dermatology, Institute for Animal Pathology, Vetsuisse Faculty of Berne, Berne, Switzerland.
authors.forename: M M
authors.initials: MM
authors.lastname: Suter
authors.affiliation: Institute of Veterinary Pathology. Electronic address: gufo@vetpath.uzh.ch.
authors.forename: F
authors.initials: F
authors.lastname: Guscetti
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D017209:Apoptosis; D002478:Cells, Cultured; D003429:Cross Reactions; D004285:Dogs; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods / Q000662:veterinary*; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology; D016159:Tumor Suppressor Protein p53 / Q000276:immunology* / Q000378:metabolism
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53