原核表达c-myc蛋白的多克隆抗体制备

探讨利用小鼠制备抗人c—myc蛋白多克隆抗体的方法。利用含PET28a／c—myc质粒的EcoliBL21，对c—myc蛋白进行原核表达，并进行纯化。WesternBlot结果只出现一条62KDa大小的特异性目的蛋白条带。用原核表达的c—myc蛋白抗原免疫5只Balb／c小鼠，最终免疫后的第7d采血，并对其血清效价进行ELISA（Enzyme—linked immunosorbnent assay）测定，抗血清达1：640000。本实验结果为c—myc单克隆抗体制备和肿瘤诊断试剂盒的研发提供基础实验数据。

Preparation of Polyclonal Antibody with Prokaryotic Expressed c-myc Protein

To explore the methods of producing antiserum with using prokaryotic expressed c - mye protein, we designed a method of immunization. The c - myc protein expressed in E. eoli strain BL21 was identified by Western blot. Five Balb/c mice were immunized with purified c - mye protein for 3times . Then the serum was picked up, and the titer of antibody was measured by ELISA （Enzyme- linked immunosorbnent assay）. The titer of antiserum was 1：64-0000. The antiserum prepared has high - titer and high - affinity,lays a foundation for further research on producing a monoclonal anti e - myc antibody.