摘要
目的 利用大肠杆菌中表达的融合蛋白GST nDOC 2 ,制备兔抗DOC 2的多克隆抗体。方法 将克隆有人DOC 2氨基端PID结构域 (暂命名为nDOC 2 )的原核表达载体pGEX 4T nDOC 2转化大肠杆菌 ,IPTG诱导表达融合蛋白GST nDOC 2 ,经包涵体洗涤后 ,用其免疫兔 ,制备兔抗人DOC 2抗血清。并以ELISA、Westernblot和免疫组化的方法对抗血清进行鉴定。结果 用ELISA法测定抗血清的效价为 1∶32 0 0 0 ;Westernblot结果显示对应于GST nDOC 2融合蛋白位置上有阳性条带 ;免疫组化结果显示人正常卵巢上皮细胞胞浆呈阳性染色 ,人卵巢浆液性囊腺瘤上皮细胞染色呈弱阳性 ,而人卵巢癌浆液性囊腺癌上皮细胞呈阴性染色。结论 成功制备了兔抗人DOC 2抗血清 ,为研究DOC
Objective To prepare antiserum against human DOC-2 using fusion protein GST-nDOC-2. Methods E.coli DH5α transformed with the recombinant plasmid pGEX-4T-nDOC-2 was induced by IPTG to express fusion protein GST- nDOC-2. Antiserum against hDOC-2 was prepared by immunizing rabbit with the fusion protein GST- nDOC-2. The antiserum was identified by ELISA, Western blot, and immunohistochemical staining. Results The antiserum titer was 1∶32 000. Western blotting showed a positive band. The hDOC-2 was detected in normal human ovarian tissue and ovarian serous cystadenoma, but not in ovarian serous cystadenocarcinoma. Conclusion The rabbit antiserum against hDOC-2 is prepared successfully.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2004年第6期465-467,471,共4页
Immunological Journal
基金
陕西省自然科学基金资助项目 (2 0 0 2C2 1 2 )
