Identification of human myocardial proteins separated by two‐dimensional electrophoresis using an effective sample preparation for mass spectrometry
- 1 January 1996
- journal article
- two dimensional-electrophoresis
- Published by Wiley in Electrophoresis
- Vol. 17 (10), 1643-1650
- https://doi.org/10.1002/elps.1150171027
Abstract
Peptide mass fingerprinting is a powerful tool for the identification of proteins separated by two‐dimensional gel electrophoresis (2‐DE). The identification of in‐gel digested proteins by peptide mass fingerprinting was significantly improved in comparison to blot‐digests by using a peptide‐collecting device. This device allows the effective purification and concentration of enzymatic digests of low‐intensity spots without expensive equipment and is described in detail. Sensitivity in the fmol range was demonstrated by unequivocal identification of bovine serum albumin after sodium dodecyl sulfate — polyacrylamide gel electrophoresis. Furthermore the high performance liquid chromatography pattern of in‐gel digests indicated a 2‐ to 3‐fold higher yield of the separated peptides. Therefore, a higher amount of the peptides was available to perform N‐terminal sequencing. The identification of 16 proteins from a high‐resolution 2‐DE gel map of human myocardium tissue has been achieved by means of this technique. Three of these proteins were associated with changes in spot intensity with dilated cardiomyopathy.Keywords
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