title: Characterization of a new monoclonal antibody against PAX5/BASP in 1525 paraffin-embedded human and animal tissue samples.
abstract: BACKGROUND
  We describe the newly generated DAK-PAX5 monoclonal antibody raised against a fixation-resistant epitope of the human PAX5/BSAP molecule.
  
  
  METHODS
  Following Western-blot, absorption, and chess-board titration tests, and optimization of antigen-retrieval and detection methods, DAK-Pax5 was used in parallel with a reference antibody (clone 24) on tissue micro-arrays (TMAs) constructed from normal human and animal tissues and from hematologic and nonhematologic human malignancies. Such TMAs were also tested with an anti-PAX2 antibody.
  
  
  RESULTS
  DAK-Pax5 reacted with normal human and animal B-cells and with 460/473 B-cell non-Hodgkin lymphomas (B-NHLs). All plasmacytomas/plasmablastic tumors (n=13) and T/NK-cell neoplasms (n=264) turned out consistently negative as did acute myelogenous leukaemias (n=19) except 2 carrying t(8;21). Positivity was found in 6/6 and 155/169 lymphocyte predominant and classical HLs, respectively, although the staining intensity varied through cases. Among 521 nonhematologic malignancies, DAK-Pax5 reacted with 22/399 carcinomas (4/11 neuroendocrine, 2/4 Merkel-cell, 4/21 prostatic, 1/11 urothelial, 1/26 renal, 2/12 cervical squamous-cell, 3/13 ovarian, and 5/75 colonic). When compared with clone 24, DAK-Pax5 produced a stronger positivity in most if not all B-NHLs and HLs. No cross-reactivity with the anti-PAX2 antibody was recorded.
  
  
  CONCLUSIONS
  DAK-Pax5 represents a new reliable tool for diagnostics and research.
authors.affiliation: Departments of Haematology and Clinical Oncology L. and A. Seràgnoli, Pathology and Haematopathology Unit, St. Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 9, Bologna, Italy. claudioagostinelli@gmail.com
authors.forename: Claudio
authors.initials: C
authors.lastname: Agostinelli
authors.forename: Elena
authors.initials: E
authors.lastname: Sabattini
authors.forename: Jakob Oemar
authors.initials: JO
authors.lastname: Gjørret
authors.forename: Simona
authors.initials: S
authors.lastname: Righi
authors.forename: Maura
authors.initials: M
authors.lastname: Rossi
authors.forename: Manuela
authors.initials: M
authors.lastname: Mancini
authors.forename: Pier Paolo
authors.initials: PP
authors.lastname: Piccaluga
authors.forename: Francesco
authors.initials: F
authors.lastname: Bacci
authors.forename: Teresa
authors.initials: T
authors.lastname: Marafioti
authors.forename: Giuliano
authors.initials: G
authors.lastname: Bettini
authors.forename: Brunangelo
authors.initials: B
authors.lastname: Falini
authors.forename: Stefano A
authors.initials: SA
authors.lastname: Pileri
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001402:B-Lymphocytes / Q000737:chemistry* / Q000473:pathology; D014408:Biomarkers, Tumor / Q000737:chemistry* / Q000276:immunology*; D015153:Blotting, Western; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008228:Lymphoma, Non-Hodgkin / Q000276:immunology / Q000473:pathology; D008297:Male; D051379:Mice; D009369:Neoplasms / Q000737:chemistry* / Q000175:diagnosis / Q000473:pathology* / Q000648:ultrastructure; D051757:PAX5 Transcription Factor / Q000737:chemistry / Q000276:immunology; D016612:Paraffin Embedding / Q000379:methods; D040081:Protein Array Analysis
chemical_list: D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D000939:Epitopes; D051757:PAX5 Transcription Factor; C497281:PAX5 protein, human