[Preparation and characterization of monoclonal antibodies against rat Nogo molecule].
OBJECTIVE
To prepare monoclonal antibodies against rat Nogo molecule and study the distribution of Nogo in rat central nervous system (CNS).


METHODS
Murine mAbs were prepared by hybridoma technique. The distribution of Nogo in rat CNS was identified by immunofluorescent histochemical staining.


RESULTS
Three hybridoma cell lines, FMU-Nogo1, FMU-Nogo2 and FMU-Nogo3, secreting mAbs against rat Nogo molecule were obtained. The titers of ascitic mAbs reached to 10(-6) and the Ig subclass of FMU-Nogo1, FMU-Nogo2 and FMU-Nogo3 was IgG2b(kappa), IgG1(kappa) and IgG1(kappa), respectively. Moreover, these mAbs could be used in immunofluoresent histochemical staining. The results showed that rat Nogo molecule could be detected in Purkinje cell layer and granual layer of rat cerebellum and on oligodendrocyte in white matter of rat spinal cord.Furthermore, these mAbs could be used in Western blot for detecting Nogo molecule present in rat spinal cord.


CONCLUSIONS
Three mAbs against rat Nogo molecule were successfully prepared, which can provide a useful tool in research on the structure and functions of Nogo molecule in CNS.
Song
Chao-jun
CJ
Department of Immunology,Fourth Military Medical University,Xi'an 710032, China.
Cheng
Xi-ping
XP
Liu
Xue-song
XS
Zhu
Yong
Y
Zhang
Ping
P
Xu
Xiao-guang
XG
Liu
Ying
Y
Jin
Bo-quan
BQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D005260:Female; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009185:Myelin Proteins / Q000032:analysis / Q000276:immunology*; D000070798:Nogo Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000737:chemistry
D000911:Antibodies, Monoclonal; D009185:Myelin Proteins; D000070798:Nogo Proteins; C000603201:Rtn4 protein, mouse; C000603197:Rtn4 protein, rat