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目的:制备CD151-整合素α6β1结合域的多克隆抗体,探讨其免疫学特性。方法:化学合成CD151-整合素α6β1结合域的多肽CGQRDHASNIYKVEG-KLH,以此多肽免疫BALB/c小鼠,制备多克隆抗体。采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)测定该抗体的效价,采用Western blotting、细胞免疫荧光染色、免疫组织化学染色的方法鉴定抗体+++。结果:成功获得抗CD151-整合素α6β1结合域的多克隆抗体,ELISA试验显示该抗体效价为1:64000,Western blotting结果显示该抗体有较好的特异性,细胞免疫荧光染色和免疫组织化学染色显示阳性染色定位于肝癌细胞膜和细胞浆。结论:制备的抗CD151-整合素α6β1结合域的多克隆抗体具有较高的效价和特异性,可用于Western blotting、细胞免疫荧光染色和免疫组织化学染色等试验,该多克隆抗体对肝癌抗复发的治疗具有重要意义,值得进一步研究。
OBJECTIVE: To prepare polyclonal antibodies against CD151-integrin α6β1 binding domain and investigate its immunological properties. Methods: Polyclonal antibody was prepared by immunization of BALB / c mice with CGQRDHASNIYKVEG-KLH, a CD151-integrin α6β1 binding domain. The antibody titer was determined by enzyme-linked immunosorbent assay (ELISA), and the antibody +++ was identified by Western blotting, immunofluorescence and immunohistochemical staining. Results: The anti-CD151-integrin α6β1 binding domain polyclonal antibody was successfully obtained. The ELISA titer of this antibody was 1: 64000. Western blotting showed that the antibody had good specificity. Immunofluorescence staining and immunohistochemistry Staining showed that the positive staining located in the liver cancer cell membrane and cytoplasm. CONCLUSION: The prepared polyclonal antibody against CD151-integrin α6β1 binding domain has high potency and specificity and can be used for the experiments such as Western blotting, immunofluorescence staining and immunohistochemical staining. The polyclonal antibody against hepatocarcinoma The treatment of recurrence is of great significance and deserves further study.