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首页> 外文期刊>Signal transduction: Receptors, mediators and genes: Official journal of the Signal Transduction Society >Double staining of proteins after separation in SDS gels with Ruthenium Bathophenantroline Disulfonate and Silver is compatible with MALDI-TOF mass spectrometry
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Double staining of proteins after separation in SDS gels with Ruthenium Bathophenantroline Disulfonate and Silver is compatible with MALDI-TOF mass spectrometry

机译：钌菲咯啉二磺酸钌和银在SDS凝胶中分离后蛋白质的双重染色与MALDI-TOF质谱兼容

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摘要

Sensitive peptide mass spectrometry techniques are increasingly used to identify molecular components of intracellular signaling networks. The detection of low abundant, electrophoretically separated proteins requires sensitive SDS gel protein stains, such as silver or fluorescent Ruthenium dyes. Both silver or Ruthenium based fluorescent dyes such as Ruthenium II Tris bathophenanthroline disulfonate (RuBPS) have unique advantages and disadvantages: some proteins stain better with silver whereas others stain better with Ruthenium dyes. Here, we demonstrate that RuBPS-stained stained proteins after separation in SDS gels can be re-stained with silver and that proteins from double-stained gels can still be subjected to MALDI-TOF mass spectrometry without adverse effects. Thus, our new double-staining technique not only produces a permanent record of RuBPS-stained gels, but more importantly, offers also an optimal staining procedure for electrophoretically separated proteins.
机译：灵敏的肽质谱技术越来越多地用于鉴定细胞内信号传导网络的分子成分。电泳分离的低丰度蛋白质的检测需要灵敏的SDS凝胶蛋白质染色剂，例如银或荧光钌染料。银基或钌基荧光染料（如Ruthnium II Tris邻菲咯啉二磺酸盐（RuBPS））都有独特的优缺点：有些蛋白质对银的染色效果更好，而另一些对钌染料的染色效果更好。在这里，我们证明了在SDS凝胶中分离后RuBPS染色的染色蛋白可以用银重新染色，并且来自双重染色凝胶的蛋白仍可以进行MALDI-TOF质谱分析而没有不利影响。因此，我们的新双染色技术不仅可以永久记录RuBPS染色的凝胶，而且更重要的是，还可以为电泳分离的蛋白质提供最佳的染色程序。

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