摘要
目的制备抗人生长激素(growth hormone,GH)的多克隆抗体并鉴定其性能。方法真核细胞表达质粒pcDNA3.0-GHcDNA免疫家兔,制备rhGH多克隆抗体;ELISA法检测抗体效价。亲和层析法纯化后,进行Western blot、免疫组织化学和激光共聚焦显微镜检测,鉴定抗体的特异性与适用范围。结果得到兔抗人重组人生长激素rhGH多克隆抗体,效价达1:40000。纯化后的抗体可特异识别超表达的hGH和人内源性GH,可用于免疫组织化学实验、Western blot及激光共聚焦。结论用质粒免疫家兔制备的抗GH抗体具有较高的效价以及特异性,为hGH的功能性研究提供了有力的支持。
To prepare,purify,and characterize the polyclonal antibody against recombinant human Growth Hormone（rhGH）.The plasmid pcDNA3.0-GHcDNA was injected into New Zealand rabbits to prepare the polyclonal antibody specifically targeting rhGH.ELISA assay was used to detect the titer of the antibody.After purification by affinity chromatography,the antibody was identified by Western blot, immunohistochemical assay,and laser scanning confocal microscope（LSCM）.The titer of polyclonal antibody was 1：4×10^4.The polyclonal antibody has high specificity to the high expressed hGH and the endogenous hGH.Our work prepared specific antibody against hGH, which will be helpful for further study on hGH function.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第1期72-73,80,共3页
Immunological Journal
基金
河南省教育厅自然科学资助项目(200510472009)
