论文部分内容阅读
目的:制备METT11D1(methyltransferase 11 domaincontaining1.简称其为GA9)(GenBank:AK024512)的多克隆抗体,并对其特异性进行鉴定。方法:融合表达GST-GA9(1-228aa)和GST-GA9(229-456aa)蛋白,利用包涵体纯化蛋白方法纯化蛋白,常规免疫小鼠,制备多克隆抗体。利用转染带有FLAG标签的FLAG-GA9真核表达载体的293T细胞裂解物,Western blot检测抗体的特异性。结果:获得了GST-GA9(1-228aa)和GST-GA9(229-456aa)融合蛋白;利用这些融合蛋白得到的多克隆抗体可特异识别FLAG-GA9蛋白。结论:成功得到可特异识别GA9的多克隆抗体,为进一步研究GA9的功能奠定了坚实的基础。
OBJECTIVE: To prepare a polyclonal antibody against METT11D1 (GenBank: AK024512), and to identify its specificity. Methods: Polyclonal antibody was prepared by fusion of GST-GA9 (1-228aa) and GST-GA9 (229-456aa) proteins. The protein was purified by inclusion body purification and the mice were routinely immunized. The specificity of the antibody was detected by Western blot using 293T cell lysate transfected with the FLAG-tagged FLAG-GA9 eukaryotic expression vector. Results: GST-GA9 (1-228aa) and GST-GA9 (229-456aa) fusion proteins were obtained. The polyclonal antibodies derived from these fusion proteins could specifically recognize FLAG-GA9 protein. Conclusion: The polyclonal antibody that can specifically recognize GA9 was successfully obtained, which laid a solid foundation for further study on the function of GA9.