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目的:制备兔抗信号转导与转录激活因子3(Stat3)抗体,分析其在乳腺癌诊断中的价值.方法:应用人工重组小鼠Stat3蛋白,免疫雄性家兔制备抗Stat3抗体;用免疫印迹法分析检测抗Stat3抗体的纯度和特异性;应用免疫组化染色法检测65例乳腺癌和30例正常乳腺组织中Stat3蛋白的表达.结果:用Stat3蛋白免疫家兔被免疫6wk后,其血清中抗Stat3抗体效价>1∶256,000(A450nm=0.40);SDS-PAGE和免疫印迹分析均在分子质量(Mr)92000处有明显的带;65例乳腺癌组织的细胞浆和细胞核中,Stat3蛋白表达量在不同的组织学分级之间有统计学差异(P<0.05).结论:高效价和特异性强的抗Stat3抗体被成功制备.该抗体可用于乳腺癌组织细胞Stat3蛋白表达的检测.
OBJECTIVE: To prepare rabbit anti-STAT3 antibody and analyze its value in the diagnosis of breast cancer.Methods: The anti-Stat3 antibody was prepared by immunizing male rabbits with recombinant Stat3 protein of mice, Assay was used to detect the purity and specificity of anti-Stat3 antibody.The expression of Stat3 protein in 65 cases of breast cancer and 30 cases of normal breast tissue was detected by immunohistochemical staining.Results: After immunized with Stat3 for 6 weeks, the serum The titer of anti-Stat3 antibody was> 1:256,000 (A450nm = 0.40). SDS-PAGE and Western blotting showed obvious bands at the molecular weight of 92000. In the cytoplasm and nuclei of 65 cases of breast cancer, Stat3 The expression of Stat3 was significantly different between different histological grades (P <0.05) .Conclusion: The anti-Stat3 antibody with high titer and specificity was successfully prepared and could be used to detect the expression of Stat3 protein in breast cancer cells .