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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。
Objective To prepare polyclonal antibodies against human growth hormone (GH) and identify its properties. Methods Rabbit immunized with eukaryotic expression plasmid pcDNA3.0-GHcDNA to prepare rhGH polyclonal antibody; ELISA antibody titer. Purification by affinity chromatography, Western blot, immunohistochemistry and laser scanning confocal microscopy to identify the specificity and scope of the antibody. The results obtained rabbit anti-human recombinant human rhGH polyclonal antibody titer of 1: 40,000. The purified antibody can specifically recognize over-expressed hGH and human endogenous GH, and can be used for immunohistochemistry, Western blot and confocal laser scanning. CONCLUSION: The anti-GH antibody prepared from rabbits immunized with plasmids has high potency and specificity and provides strong support for the functional study of hGH.