title: [Preparation and application of monoclonal antibody against human vascular endothelial cadherin 5].
abstract: OBJECTIVE
  To prepare and identify monoclonal antibody (mAb) against human vascular endothelial cadherin 5 (VECDH5).
  
  
  METHODS
  BALB/c mice were immunized with recombinant VECDH5 protein for preparing mAb using hybridoma technique. The positive clones were confirmed and selected by indirect ELISA for titer determination. Western blotting, flow cytometry, immunofluorescence staining, immunohistochemical staining were performed to identify the specificity and epitope.
  
  
  RESULTS
  One hybridoma cell strain secreting specific mAb against VECDH5 was obtained (2C11). ELISA showed that the titer of the ascites was 1:10 000. Western blotting, flow cytometry, immunofluorescence, immunohistochemistry demonstrated that the mAb could specifically recognize and bind VECDH5. Epitope identification showed that the amino acid sequence recognized by 2C11 was LDREVVPWYNLTVEA.
  
  
  CONCLUSIONS
  We have prepared mAb against human VECDH5, which has good binding ability and specificity.
authors.affiliation: Clinical Laboratory, First Affiliated Hospital, Soochow University, Suzhou 215006, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Ding
authors.affiliation: Clinical Laboratory, First Affiliated Hospital, Soochow University; Jiangsu Provincial Institute of Hematology, Suzhou 215006, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: He
authors.affiliation: Cyrus Tang Hematology Research Center, Soochow University, Suzhou 215006, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Yang
authors.affiliation: Cyrus Tang Hematology Research Center, Soochow University, Suzhou 215006, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zuo
authors.affiliation: Clinical Laboratory, First Affiliated Hospital, Soochow University, Suzhou 215006, China. *Corresponding author, E-mail: ybh8527@163.com.
authors.forename: Binghua
authors.initials: B
authors.lastname: Yang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D015820:Cadherins / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry; D056945:Hep G2 Cells; D006801:Humans; D006825:Hybridomas; D008297:Male; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D000099152:Cadherin 5
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015820:Cadherins; D000939:Epitopes; D000099152:Cadherin 5