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目的：为了制备ｎＮＯＳ特异性抗体。方法：用ＰＣＲ的方法克隆出ｎＮＯＳ７０８～８８１ａａ片段的编码基因并将之插入ｐＥＴ２８ａ原核表达载体。结果：成功地在大肠杆菌中获得高效表达。经ＨｉｓＴａｇＳｅｐｈａｒｏｓｅ和电泳回收法得到纯蛋白，将此蛋白免疫兔，制备出ｎＮＯＳ特异性抗体，抗体效价可达１∶８０００。利用该抗体成功地检测了正常肌肉和ＤＭＤ肌肉中ｎＮＯＳ的含量变化，证实ＤＭＤ肌肉中ｎＮＯＳ的含量明显低于正常肌肉。结论：制备的抗体具有较高的特异性，对ｎＮＯＳ的免疫检测具有重要应用价值
Purpose: To prepare nNOS-specific antibodies. Methods: The coding gene of nNOS708 ~ 881aa fragment was cloned by PCR and inserted into pET28a prokaryotic expression vector. Results: High expression was successfully obtained in E. coli. Pure protein was obtained by HisTag-Sepharose and electrophoresis recovery method, and then the protein was immunized into rabbit to prepare nNOS-specific antibody with antibody titer up to 1: 8000. Using this antibody, we successfully detected the changes of nNOS in normal muscle and DMD muscle, and confirmed that the content of nNOS in DMD muscle was significantly lower than normal muscle. Conclusion: The prepared antibody has high specificity and is of great value to the immunoassay of nNOS