Epitope tags are short, linear antibody recognition sequences that enable detection of tagged fusion proteins by antibodies. Epitope tag position and neighboring sequences potentially affect its recognition by antibodies, and such context-dependent differences in tag binding may have a wide-ranging effect on data interpretation. We tested by Western blotting six antibodies that recognize the c-Myc epitope tag, including monoclonal antibodies 9E10, 4A6, 9B11, and 71D10 and polyclonal antibodies 9106 and A-14. All displayed context-dependent differences in their ability to detect N- or C-terminal Myc-tagged proteins. In particular, clone 9E10, the most cited Myc-tag antibody, displayed high context-dependent detection variability, whereas others, notably 4A6 and 9B11, showed much less context sensitivity in their detection of Myc-tagged proteins. The very high context sensitivity of 9E10 was further substantiated by peptide microarray analyses. We conclude that recently developed, purpose-made monoclonal antibodies specific for Myc have much more uniform reactivity in diverse assays and are much less context sensitive than is the legacy antibody 9E10.

中文翻译：

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Myc标签单克隆抗体9E10显示高度可变的表位识别，具体取决于邻近序列的背景。

表位标签是短的线性抗体识别序列，可通过抗体检测标记的融合蛋白。表位标签的位置和邻近序列可能会影响抗体对其的识别，并且标签绑定中的此类上下文相关差异可能会对数据解释产生广泛影响。我们通过蛋白质印迹测试了六种识别c-Myc表位标签的抗体，包括单克隆抗体9E10、4A6、9B11和71D10以及多克隆抗体9106和A-14。所有这些在检测N或C末端Myc标记的蛋白质的能力上均显示出上下文相关的差异。特别是，克隆9E10（引用最多的Myc-tag抗体）显示出较高的上下文相关检测变异性，而其他（尤其是4A6和9B11）在检测Myc标记的蛋白质时显示出较低的上下文敏感性。肽微阵列分析进一步证实了9E10的非常高的上下文敏感性。我们得出的结论是，与传统抗体9E10相比，最近开发的针对Myc的特制单克隆抗体在多种测定法中的反应性更高，对环境的敏感性更低。