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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Rehulková, Helena Chalupová, Jana Sebela, Marek Rehulka, Pavel |
| Description | Country affiliation: Czech Republic Author Affiliation: Rehulková H ( Faculty of Military Health Sciences, Institute of Molecular Pathology, University of Defence, Trebesská 1575, CZ-50001 Hradec Králové, Czech Republic.) |
| Abstract | Peptide samples derived from enzymatic in-gel digestion of proteins resolved by gel electrophoresis often contain high amount of salts originating from reaction and separation buffers. Different methods are used for desalting prior to matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS), e.g. reversed-phase pipette tip purification, on-target washing, adding co-matrices, etc. As a suitable matrix for MALDI MS of peptides, alpha-cyano-4-hydroxycinnamic acid (CHCA) is frequently used. Crystalline CHCA shows the ability to bind peptides on its surface and because it is almost insoluble in acidic water solutions, the on-target washing of peptide samples can significantly improve MALDI MS signals. Although the common on-target washing represents a simple, cheap and fast procedure, only a small portion of the available peptide solution is efficiently used for the subsequent MS analysis. The present approach is a combination of the on-target washing principle carried out in a narrow-end pipette tip (e.g. GELoader tip) and preconcentration of peptides from acidified solution by passing it through small CHCA crystals captured inside the tip on a glass microfiber frit. The results of MALDI MS analysis using CHCA-tip peptide preconcentration are comparable with the use of homemade POROS R2 pipette tip microcolumns. Advantages and limitations of this approach are discussed. |
| File Format | HTM / HTML |
| ISSN | 10765174 |
| Issue Number | 1 |
| Volume Number | 45 |
| e-ISSN | 10969888 |
| Journal | Journal of Mass Spectrometry |
| Language | English |
| Publisher | Wiley |
| Publisher Date | 2010-01-01 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Spectroscopy Discipline Chemistry Techniques Chemical Fractionation Methods Coumaric Acids Chemistry Peptides Spectrometry, Mass, Matrix-assisted Laser Desorption-ionization Isolation & Purification Salts Tandem Mass Spectrometry Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Medicine |
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