Abstract
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.
Highlights
A method for increasing the productivity of ESI LC-MS/MS was proposed and applied
One example is so called “Virtual 2-D Gel Electrophoresis” [4,5], where isoelectrofocusing in IPG strips is used as a first dimension, but the conventional second separation by SDS-PAGE is replaced by MALDI-MS
Usage of ESI LC-MS/MS, instead of MALDI-MS, in combination with classical electrophoretic methods allows the revelation of more information; especially when very complex samples like mammalian cell extracts are analyzed [6,7,8,9]
Summary
ARTICLE INFO Method name: A semi-virtual two dimensional gel electrophoresis: IF–ESI LC-MS/MS Keywords: Proteoforms, Large-scale, Identification, IEF, Shotgun, 2DE Article history: Received 3 February 2017; Accepted 25 August 2017; Available online 31 August 2017
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