摘要
目的:制备抗人视黄醇结合蛋白4(retinal binding protein4,RBP4)的多克隆抗血清并用于妊娠糖尿病(GDM)血液和胎盘的检测。方法:用自制rhRBP4蛋白免疫新西兰兔制备多克隆抗血清,Western blot检测特异性,ELISA法测定效价,收集20例GDM患者和15例正常孕妇的血液和胎盘,检测RBP4的水平和分布。结果:制备的抗血清效价为1∶128000。与正常孕妇相比,GDM患者血清中RBP4水平明显升高,其分娩胎盘的滋养层细胞质和胞膜中RBP4也显著高表达。结论:制备的多克隆抗血清可初步用于GDM患者血清或组织样本中RBP4的检测,为进一步用于临床检测奠定基础。
Objective To prepare the rabbit polyclonal antiserum against retinal binding protein 4 （RBP4） which could be applied to detect blood and placental in gestational diabetic mellitus （GDM）. Methods The polyelonal antiserum to rhRBP4 were prepared by immunizing New Zealand rabbits with self-made protein rhRBP4. The specificity and titer of antiserum were determined by western blot and ELISA respectively. RBP4 levels and distributions in serum and placenta of 20 GDM patients and 15 normal pregnant women were analyzed by western blot and immunohistochemistry respectively. Results The titer of polyclonal antiserum to rhRBP4 was about 1：128 000. RBP4 serum levels of GDM patients were significantly higher than normal pregnant women, mainly distributed in the cytoplasm and membrane of trophocyte. Conclusion The polyclonal antiserum were successfully prepared and could be preliminary used to detect RBP4 in serum and tissue samples of GDM patients, which laid as a foundation for further clinical detection.
出处
《实用医学杂志》
CAS
北大核心
2013年第7期1050-1052,共3页
The Journal of Practical Medicine
基金
合肥市科技局基金项目(编号:合科[2011]25号)
关键词
糖尿病
妊娠
重组人视黄醇结合蛋白4
多克隆抗血清
血清
胎盘
Diabetes, gestational
Recombinant human retinol binding protein 4
Polyclonal antiserum
Serum
Placenta
