A semi-virtual two dimensional gel electrophoresis: IF–ESI LC-MS/MS
نویسندگان
چکیده
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.
منابع مشابه
Dataset of protein species from human liver
This article contains data related to the research article entitled "Zipf׳s law in proteomics" (Naryzhny et al., 2017) [1]. The protein composition in the human liver or hepatocarcinoma (HepG2) cells extracts was estimated using a filter-aided sample preparation (FASP) protocol. The protein species/proteoform composition in the human liver was determined by two-dimensional electrophoresis (2-DE...
متن کاملConstruction of a two-dimensional gel electrophoresis protein database for the Nicotiana tabacum cv. Bright Yellow-2 cell suspension culture.
Using two-dimensional gel electrophoresis (2-DE) and electrospray-tandem mass spectrometry (ESI-MS/MS), we have started the proteome analysis of the cell line Nicotiana tabacum cv. Bright Yellow-2 (tobacco BY-2). The BY-2 cell suspension culture is widely used as a model system to study the growth and development of plant cells. We present a protocol describing the sample preparation and 2-DE, ...
متن کاملProteomics of the post-translational modifi cations: the knowns and the unknowns
Abbreviations: PTMs=post-translational modifi cations; PPIs=protein-protein interactions; SDS-PAGE= sodium dodecyl sulfate-polyacrylamide gel electrophoresis; 2D-PAGE=two-dimensional polyacrylamide gel electrophoresis; MS=mass spectrometry; ESI=electrospray ionization; MALDI=Matrix Assisted Laser Desorption Ionization; ESI-MS=electrospray ionization mass spectrometry; MALDI-MS=MALDI mass spectr...
متن کاملTowards the Full Realization of 2DE Power
Here, approaches that allow disclosure of the information hidden inside and outside of two-dimensional gel electrophoresis (2DE) are described. Experimental identification methods, such as mass spectrometry of high resolution and sensitivity (MALDI-TOF MS and ESI LC-MS/MS) and immunodetection (Western and Far-Western) in combination with bioinformatics (collection of all information about prote...
متن کاملSeparation and identification of soybean leaf proteins by two-dimensional gel electrophoresis and mass spectrometry.
To establish a proteomic reference map for soybean leaves, we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Tryptic digests of 260 spots were subjected to peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS. Fifty-three of these protein spots were id...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 4 شماره
صفحات -
تاریخ انتشار 2017