摘要
目的制备胃动素受体多克隆抗体和研究该受体在人胃中的组织定位。方法通过合成肽耦联钥孔戚血蓝蛋白(keyholelimpethemocyanin,KLH)制备抗体,以ELISA方法测定效价。用含有GPR38基因(胃动素受体)的质粒转染293细胞株,以RT-PCR方法鉴定。Westernblot和免疫组织化学法检测转染的293细胞和组织中胃动素受体的表达。结果制备的兔抗人的多抗效价达1∶500000,有较好的特异性。成功转染293细胞,该细胞能够瞬时表达胃动素受体,而该受体在人胃窦胃体的黏膜腺上皮中表达。结论应用合成肽-KLH成功地制备了胃动素受体的兔抗人多克隆抗体,用该抗体检测到人胃窦胃体黏膜上皮细胞存在胃动素受体的表达。
Objective To raise polyclonal antibody against motilin receptor and investigate the expression of this receptor in human stomach. Methods Synthesized peptide was used to raise rabbit anti human antibody against N terminal of motilin receptor and ELISA method was used to measure the titer of the antiserum. Electro transfect 293 cells were transfected with GPR38 plasmid and RT PCR was used to confirm successful transfection. Western blot and immunohistochemistry method were applied to study protein expression of motilin receptor in transfected 293 cells and human stomach tissues. Results The titer of raised antibody was 1∶500 000 with good specificity. GPR38 plasmid was successfully transfected in 293 cells with protein expression. Motilin receptor was shown to express in glandular epithelium of antrum and corpus of stomach. Conclusions Polyclonal antibody against motilin receptor is successfully raised by synthesized peptide with high titer and good specificity. Motilin receptor expresses in glandular epithelial cells of antrum and corpus of human stomach.
出处
《中华胃肠外科杂志》
CAS
2002年第4期252-255,共4页
Chinese Journal of Gastrointestinal Surgery
基金
国家自然科学基金资助项目(30070738)
