J. Craig Venter Institute, Rockville, MD, USA.
Methods Mol Biol. 2021;2261:25-34. doi: 10.1007/978-1-0716-1186-9_3.
Effective and reliable protease digestion of biological samples is critical to the success in bottom-up proteomics analysis. Various filter-based approaches using different types of membranes have been developed in the past several years and largely implemented in sample preparations for modern proteomics. However, these approaches rely heavily on commercial filter products, which are not only costly but also limited in membrane options. Here, we present a plug-and-play device for filter assembly and protease digestion. The device can accommodate a variety of membrane types, can be packed in-house with minimal difficulty, and is extremely cost-effective and reliable. Our protocol offers a versatile platform for general proteome analyses and clinical mass spectrometry.
有效的和可靠的蛋白酶消化生物样本是至关重要的成功在自上而下的蛋白质组学分析。各种基于过滤的方法使用不同类型的膜已经开发了在过去的几年中,并在样品制备中得到了广泛的应用现代蛋白质组学。然而,这些方法严重依赖于商业过滤产品,这不仅是昂贵的,而且还局限于膜的选择。在这里,我们提出了一个即插即用的设备进行过滤组件和蛋白酶消化。该设备可以容纳各种类型的膜,可以包装在内部最小的困难,和是非常经济高效和可靠的。我们的协议提供了一个通用的平台进行一般的蛋白质组分析和临床质谱。
