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Objective To prepare polyclonal antibodies of anti Nogo-66, the extracellular region of one central nervous system neurite regeneration inhibitor Nogo, which could be used to further identification and functional study of Nogo molecule.Methods Preparing rabbit anti rat Nogo-66 polyclonal antibodies with a purified Nogo-66 fusion protein expressed in E.coli system. Studying its specificity by Western-blot and immuno-histochemical techniques and identifying its biological activity in PC12 cells.Results The high titer (1∶[KG-*2]10 000) anti rat Nogo-66 polyclonal antibodies were obtained.This antibody could specifically recognize the Nogo protein expressed in E.coli system.Immuno-histochemical staining indicated that the Nogo was widely expressed in rat spinal cord neurons and oligodendrocytes.It could effectively block the neurite extensioninhibition of Nogo protein in PC12.Conclusion Successful preparation of anti rat Nogo polyclonal antibodies provides a useful tool in identification or further functional study of Nogo molecule.
RESUMO
Objective To block the bio function of TGF ? by the application of anti TGF ? polyclonal antibody, and observe its efficacy of prevention of abdominal adhesion. Methods The postoperative abdominal adhesion model was established in SD rats. The drugs were administrated by abdominal injection with saline (control group), sodium hyaluronate( HA group), and varied dosage of anti TGF? (anti TGF? groups) respectively. The adhesion was scored 21days later, while 30 of them were executed on the day 3 and day 10 after operation respectively. The expression of TGF ? was checked by immunohistochemistry in the samples obtained from the adhesion sites. Results The score of adhesion in anti TGF ? group (2.4?0.99) was significantly lower than that in control group (6.0?1.25) and HA group (3.4?1.03); in different dosage of anti TGF, the 50?g group showed its economical efficacy; in the control and HA groups the expression of TGF ? had a time dependent manner, which reachs to maximum in the day 3, and could be reduced by antibody. Conclusions The polyclonal antibody of TGF ? shows the power to prevent the postoperative abdominal adhesion in animal model, the mechanism of which is due to inhibition of TGF ? expression.