一种半虚拟二维凝胶电泳：免疫荧光-电喷雾电离液相色谱-串联质谱法
A semi-virtual two dimensional gel electrophoresis: IF-ESI LC-MS/MS.
作者信息
Naryzhny Stanislav, Zgoda Victor, Kopylov Artur, Petrenko Elena, Archakov Аlexander
机构信息
Orekhovich Institute of Biomedical Chemistry of Russian Academy of Medical Sciences, Pogodinskaya 10, Moscow, 119121, Russia.
B.P. Konstantinov Petersburg Nuclear Physics Institute, National Research Center "Kurchatov Institute", Orlova roscha, Gatchina, Leningrad region, 188300, Russia.
出版信息
MethodsX. 2017 Aug 31;4:260-264. doi: 10.1016/j.mex.2017.08.004. eCollection 2017.
A method for increasing the productivity of ESI LC-MS/MS (electrospray ionization-liquid chromatography-tandem mass spectrometry) was proposed and applied. After IF (isoelectric focusing) of the sample using IPG (immobilized pH gradient) strip, the strip was cut to sections, and every section was treated according to trypsinolysis protocol for MS/MS analysis. The peptides produced were further analyzed by ESI LC-MS/MS. The procedure allows to: •identify many more proteins and proteoforms compared to shotgun analysis of extracts.•build a semi-virtual 2DE map of identified proteins.
提出并应用了一种提高电喷雾电离-液相色谱-串联质谱（ESI LC-MS/MS）生产率的方法。使用固定化pH梯度（IPG）条对样品进行等电聚焦（IF）后，将条带切成小段，每段按照胰蛋白酶消化方案处理以进行串联质谱分析。产生的肽段通过ESI LC-MS/MS进一步分析。该程序能够：•与提取物的鸟枪法分析相比，鉴定出更多的蛋白质和蛋白质变体。•构建已鉴定蛋白质的半虚拟二维电泳图谱。
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