title: [Preparation and identification of the polyclonal antibody against human VSTM1].
abstract: OBJECTIVE
  To prepare and characterize the polyclonal antibody against human VSTM1.
  
  
  METHODS
  VSTM1 has two main isoforms,  VSTM1-v1,  a type I transmembrane protein,  and VSTM1-v2,  a classical secretory protein,  lacking only the transmembrane domain compared with VSTM1-v1. Two recombinant prokaryotic proteins of VSTM1-v2,  Trx-His-S-VSTM1-v2 and GST-VSTM1-v2,  were constructed,  expressed,  purified,  and then used for immunization of New Zealand rabbits to prepare anti-VSTM1 antibody and coupling with CNBr-activated Sepharose 4B to purify the antibody by immunoaffinity chromatography,  respectively. ELISA   was performed to detect the titers of the antiserums. After purification,  the antibody was identified by Western blotting  and  immunofluorescence  cytochemistry.
  
  
  RESULTS
  The titers of the antiserums from the two immunized rabbits were both 1:10(6);. Western blotting  confirmed that the purified antibody could recognize both the overexpressed and endogenous VSTM1 specifically. Immunofluorescence cytochemistry verified that the antibody could also recognize the overexpressed VSTM1-v1 on the surface of HEK293T cells.
  
  
  CONCLUSIONS
  The rabbit antibody against human VSTM1 of a high titer has been obtained,  which can be used for recognizing endogenous VSTM1 in immunofluorescence cytochemistry.
authors.affiliation: Department of Immunology,  School of Basic Medical Sciences,  Key Laboratory of Medical Immunology,  Ministry of Health,  Center for Human Disease Genomics,  Peking University,  Beijing 100191,  China.
authors.forename: Ting
authors.initials: T
authors.lastname: Li
authors.forename: Xiao-Huan
authors.initials: XH
authors.lastname: Guo
authors.forename: Ping-Zhang
authors.initials: PZ
authors.lastname: Wang
authors.forename: Quan-Sheng
authors.initials: QS
authors.lastname: Song
authors.forename: Da-Long
authors.initials: DL
authors.lastname: Ma
authors.forename: Wen-Ling
authors.initials: WL
authors.lastname: Han
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D000918:Antibody Specificity; D003001:Cloning, Molecular; D057809:HEK293 Cells; D006801:Humans; D007114:Immunization; D008297:Male; D011817:Rabbits; D011971:Receptors, Immunologic / Q000032:analysis* / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; D011971:Receptors, Immunologic; C550469:VSTM1 protein, human