title: [Preparation and characterization of monoclonal antibody against human LSECtin].
abstract: OBJECTIVE
  To prepare and characterize monoclonal antibody (mAb) against human LSECtin (liver and lymph node sinusoidal endothelial cell C-type lectin) protein.
  
  
  METHODS
  BALB/c mice were immunized with prokaryotically expressed human LSECtin protein. The splenocytes from the immunized mice were fused with murine myeloma cells (Sp2/0) and then the mAb-positive hybridoma cells were screened by indirect ELISA. Reaction of mAb to LSECtin antigen was characterized by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM.
  
  
  RESULTS
  Eight hybridoma cells secreting mAbs were established. The isotypes of the mAbs were IgG. Ascites titers were between 1:10(6) - 1:10(7). All the mAbs recognized human LSECtin protein on LSECtin-transfected 3T3 cells and six of the mAbs specifically recognized liver sinusoidal endothelial cells.
  
  
  CONCLUSIONS
  Eight anti-LSECtin mAbs have been obtained. The characterization of the mAbs indicate that they show fine specificity by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM, which can provide a powerful reagent for the functional study of LSECtin.
authors.affiliation: Department of Immunology, Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Xue-mei
authors.initials: XM
authors.lastname: Du
authors.forename: Li
authors.initials: L
authors.lastname: Tang
authors.forename: Xiao-lan
authors.initials: XL
authors.lastname: Liu
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Jian-en
authors.initials: JE
authors.lastname: Gao
authors.forename: Fu-chu
authors.initials: FC
authors.lastname: He
authors.forename: Qi-hong
authors.initials: QH
authors.lastname: Sun
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007132:Immunoglobulin Isotypes / Q000032:analysis / Q000276:immunology; D037181:Lectins, C-Type / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; C505228:CLEC4G protein, human; D007132:Immunoglobulin Isotypes; D037181:Lectins, C-Type