CD133多克隆抗体制备及肿瘤干细胞鉴定

The CD133 Polyclonal Antibody Generation and Cancer Stem Cells Indentification

目的制备肿瘤干细胞标志性蛋白CD133的多克隆抗体,为进一步研究肿瘤组织中肿瘤干细胞的生物学特性以及筛选肿瘤干细胞、制备肿瘤干细胞的小鼠模型奠定基础。方法以人CD133蛋白细胞外膜表面肽段为抗原,免疫新西兰大白兔制备多克隆抗体,进而用其对肿瘤组织进行Western blot以及免疫组化染色。结果 Western blot证实该抗体可以识别过表达的myc-CD133以及内源性CD133,进一步免疫组化结果显示该抗体可以识别恶性胶质瘤中的肿瘤干细胞。结论成功制备高效、特异的CD133多克隆抗体,该抗体可以用于肿瘤组织的免疫染色及肿瘤干细胞的筛选。

Objective To generate the cancer stem cells（CSCs） specific protein CD133 polyclonal antibody for the study of the biological characteristics of CSCs in tumor tissues and CSCs screening for the mouse model.Methods The extracellular peptide of the human CD133 was injected into rabbits to generate polyclonal antibody which was used for glioblastoma（GBM） Western blot and immunohistochemistry.Results The CD133 antiserum we made could detect both overexpressed myc-CD133 and endogenous CD133 efficiently by Western blot.Immunohistochemistry indicated that the CD133 polyclonal antibody can label CSCs in GBM sections.Conlusion High efficient and specific CD133 antibody was generated successfully and could be used to label CSCs in tumor sections and screen CSCs for the mouse model.