Aim: 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.
Conclusion: 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.