[Preparations of the specific antibodies against exon 2 and exon 3 of human tau protein].
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.
Chen
Cao
C
College of Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161, China.
Wang
Gui-rong
GR
Shi
Qi
Q
Zhang
Bao-yun
BY
Mei
Guo-yong
GY
Li
Yuan
Y
Wang
Xin
X
Zhou
Rui-min
RM
Han
Jun
J
Zhao
Yu-jun
YJ
Dong
Xiao-ping
XP
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
D000906:Antibodies; D011993:Recombinant Fusion Proteins; D016875:tau Proteins