title: [Expression, purification and polyclonal antibody preparation of GST-Ccd1 fusion protein].
abstract: OBJECTIVE
  By using of Escherichia coli DH5alpha to express GST-Ccd1 fusion protein and which was purified by affinity chromatographic separation. The purified target protein was used to immunize rabbits to prepare polyclonal antibody.
  
  
  METHODS
  The previously constructed recombinant prokaryotic expression vector pGEX-5X-1-Ccd1 was transformed into Escherichia coli DH5alpha and was induced to expression by IPTG. The recombinant target protein was expressed with soluble state in Escherichia coli expression system which was separated and purified by chromatographic column stuffed with glutathione Sepharose 4B. The prepared antigen was used to immunize rabbits to get anti-Ccd1 specific rabbit original polyclonal antibody.
  
  
  RESULTS
  ELISA data demonstrated that the antibody titer of the serum was up to 1:40 000. Immunohistochemistry analysis indicated that the "home-made" antibody had a specific interaction with Ccd1 protein and which could be used for extended experimental research.
  
  
  CONCLUSIONS
  The anti-Ccd1 polyclonal antibody we had prepared had a high quality of potency and specificity. The antibody was demonstrated by experiments that which could totally fulfill the requirement of immunoblotting and immunohistochemistry study of Ccd1. The antibody provided an useful experimental tool to profoundly research the tissue expression profile, intercellular location and biological function of Ccd1.
authors.affiliation: Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wu
authors.forename: Xiao-Tang
authors.initials: XT
authors.lastname: Jing
authors.forename: Xin
authors.initials: X
authors.lastname: Ma
authors.forename: Wen-Hong
authors.initials: WH
authors.lastname: Fan
authors.forename: Ming
authors.initials: M
authors.lastname: Fan
mesh_terms: D000906:Antibodies / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000906:Antibodies; D047908:Intracellular Signaling Peptides and Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase