Yeung Yee-Guide, Stanley E Richard
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York, USA.
Curr Protoc Protein Sci. 2010 Feb;Chapter 16:Unit 16.12. doi: 10.1002/0471140864.ps1612s59.
Detergents are required for the extraction of hydrophobic proteins and for the maintenance of their solubility in solution. However, the presence of detergents in the peptide samples severely suppresses ionization in mass spectrometry (MS) analysis and decreases chromatographic resolution in LC-MS. Thus, detergents must be removed for sensitive detection of peptides by MS. This unit describes a rapid protocol in which ethyl acetate extraction is used to remove octylglucoside from protease digests without loss of peptides. This procedure can also be used to reduce interference by sodium dodecyl sulfate, Nonidet P-40, or Triton X-100 in peptide samples for MS analysis.
提取疏水性蛋白质并维持其在溶液中的溶解度需要使用去污剂。然而,肽样品中去污剂的存在会严重抑制质谱(MS)分析中的离子化,并降低液相色谱-质谱联用(LC-MS)中的色谱分辨率。因此,为了通过MS对肽进行灵敏检测,必须去除去污剂。本单元介绍了一种快速方法,其中使用乙酸乙酯萃取从蛋白酶消化物中去除辛基葡糖苷,而不会损失肽。此方法也可用于减少十二烷基硫酸钠、Nonidet P-40或Triton X-100对用于MS分析的肽样品的干扰。