title: [Preparations of the specific antibodies against exon 2 and exon 3 of human tau protein].
abstract: OBJECTIVE
  To prepare the specific antibodies against exon 2 and exon 3 of human tau protein.
  
  
  METHODS
  Sequences encoding exon 2 and exon 3 of human tau protein were amplified from human peripheral blood DNA and cloned into a prokaryotic expression vector pGEX-2T. Fusion proteins GST-E2 and GST-E3 were expressed and purified from E. coli system. The antisera were elicited by immunization of the purified fusion proteins to rabbits and mice. The specific antibodies were purified by Protein G/A and CNBr-activated sepharose 4B coupled with GST protein. The specificity and sensitivity of the purified antibodies were evaluated by Western blotting and ELISA.
  
  
  RESULTS
  Recombinant fusion proteins GST-E2 and GST-E3 were expressed and purified from E. coli, which showed Mr. 29 x 10(3). Various antisera were collected from the immunized experimental animals. Reliable immunoreactive specificity and titers of the purified antibodies against exon 2 and exon 3 of human tau protein were confirmed by Western blotting and ELISA after serial purification processes.
  
  
  CONCLUSIONS
  Four specific antibodies against exon 2 and exon 3 of human tau protein have been successfully prepared, which provides basis for analyzing the role of tau in neurodegenerative diseases.
authors.affiliation: College of Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161, China.
authors.forename: Cao
authors.initials: C
authors.lastname: Chen
authors.forename: Gui-rong
authors.initials: GR
authors.lastname: Wang
authors.forename: Qi
authors.initials: Q
authors.lastname: Shi
authors.forename: Bao-yun
authors.initials: BY
authors.lastname: Zhang
authors.forename: Guo-yong
authors.initials: GY
authors.lastname: Mei
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Rui-min
authors.initials: RM
authors.lastname: Zhou
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.forename: Yu-jun
authors.initials: YJ
authors.lastname: Zhao
authors.forename: Xiao-ping
authors.initials: XP
authors.lastname: Dong
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005091:Exons; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016875:tau Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000906:Antibodies; D011993:Recombinant Fusion Proteins; D016875:tau Proteins