title: [Preparation and characterization of a directly labeled mouse anti-human CD14 monoclonal antibody ZCH-2F9-FITC].
abstract: To prepare fluorescein isothiocyanate (FITC) directly conjugated to monoclonal antibody (McAb) anti-human CD14, ZCH-7-2F9 (2F9-FITC).
  
  
  
  After generation and purification, the purity and the murine immunoglobulin subtype of the antibody were evaluated with SDS-PAGE and multicolor flow cytometry (FCM). 2F9 McAb was directly labeled with FITC through modified Marsshall's method and the positive rate of the 2F9-FITC on different types of leukemic cells were compared with the standard CD14-FITC by FCM.
  
  
  
  A large quantity of purified 2F9 McAb was prepared. The subtype of 2F9 was murine IgG1kappa. 2F9-FITC was successfully manufactured with A295/A280 ratio of 0.44. The positive cell percentages of 2F9-FITC and CD14-FITC on the monocytes were 84.50% and 90.08%, respectively, while those on lymphocytes were only 0.52% and 1.01%. There was no significant difference between the CD14 expressions with 2F9-FITC and CD14-FITC on each type of leukemia (n=23, t=0.922, P=0.367).
  
  
  
  2F9-FITC has been successfully prepared and it can be applied in diagnosis and differentiation of monoblastic leukemias.
authors.affiliation: Department of Hematology and Oncology, The Affiliated Children's HospitalìCollege of Medicine, Zhejiang University, Hangzhou 310003, China.
authors.forename: Bo-tao
authors.initials: BT
authors.lastname: Ning
authors.forename: Yong-min
authors.initials: YM
authors.lastname: Tang
authors.forename: Hong-qiang
authors.initials: HQ
authors.lastname: Shen
authors.forename: Shi-long
authors.initials: SL
authors.lastname: Yang
authors.forename: Ying-hu
authors.initials: YH
authors.lastname: Chen
authors.forename: Hua
authors.initials: H
authors.lastname: Song
authors.forename: Shu-wen
authors.initials: SW
authors.lastname: Shi
authors.forename: Bai-qin
authors.initials: BQ
authors.lastname: Qian
authors.forename: Wei-qun
authors.initials: WQ
authors.lastname: Xu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002478:Cells, Cultured; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000032:analysis / Q000138:chemical synthesis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007945:Leukemia, Lymphoid / Q000473:pathology; D015470:Leukemia, Myeloid, Acute / Q000473:pathology; D018950:Lipopolysaccharide Receptors / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009000:Monocytes / Q000166:cytology
chemical_list: D000911:Antibodies, Monoclonal; D018950:Lipopolysaccharide Receptors; D016650:Fluorescein-5-isothiocyanate