摘要
目的 制备cbfa1多克隆抗体并进行鉴定和效价分析 ,观察其在骨和牙胚中的表达情况。方法 用自制的融合蛋白免疫新西兰大白兔 ,制备多抗血清 ,提取小鼠牙胚、颅骨和肝脏组织的总蛋白进行westernblot ,同时采用免疫组化染色观察其组织表达特性。结果 cbfa1蛋白在小鼠颅骨和牙胚组织中均有表达 ,在人骨肉瘤细胞系中呈强阳性表达。在免疫组化和westernblot的有效作用滴度分别为 1:40 0和 1:10 0 0。
Objective To prepare the cbfa1 antibody and observe the expression of it in mouse bones and tooth germs. Methods Complete Freund`s adjuvant was used when immunized the rabbit with fusion cbfa1 proteins. The specific IgG antibody was feuther purified with sulfuric acetate. Total proteins isolated from mouse tooth germs, liver and skull tissues of mouse different developing stages were used in Western blotting study. Meanwhile, immunohistochemical techniques were also applied to examine the antibody. Results cbfa 1 proteins were expressed in mouse skulls and tooth germs, not in mouse livers. All experiment above showed the antibody we prepared was correct and specific to cbfa1 proteins and the effective deepness was 1:400 in immunohistochemistry and 1∶1000 in Western blot.Conclusion We prepare the cbfa1 antibody successfully.
出处
《临床口腔医学杂志》
2003年第5期259-261,共3页
Journal of Clinical Stomatology
基金
国家自然科学基金资助项目 (30 2 71 4 1 8)
陕西省卫生厅科研基金项目 (0 2D48)
