Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT)

Watt SA, Patschkowski T, Kalinowski J, Niehaus K (2003)
Journal of Biotechnology 106(2-3): 287-300.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
The chemically-coded affinity tag (CCAT) method combines standard electrophoresis protocols with MALDI-TOF-MS analysis to identify and quantify protein abundances in complex samples in one step. This method is designed to fit into the workflow of SDS-PAGE or two-dimensional electrophoresis (2-DE) only requiring basic proteome laboratory equipment. Prior to electrophoresis two protein samples are separately labelled with a heavy or a light version of the CCAT reagent via reduced cysteines in the proteins. Equal amounts are then combined and electrophoretically separated. Proteins can then be excised from the gel to obtain their peptide mass fingerprint by mass spectrometry. This fingerprint enabled not only identification, but also quantification by comparing relative peak intensities of CCAT-labelled peptides. In this article, we display how the CCAT method can be used to analyse two protein samples in one gel and that the peak intensities of labelled peptides reflect the abundance of a protein in it. (C) 2003 Published by Elsevier B.V.
Stichworte
affinity; two-dimensional gel electrophoresis; quantitative proteomics; tags; biotin-PEO3-iodoacetamide; CCAT
Erscheinungsjahr
2003
Zeitschriftentitel
Journal of Biotechnology
Band
106
Ausgabe
2-3
Seite(n)
287-300
ISSN
0168-1656
Page URI
https://pub.uni-bielefeld.de/record/1609350

Zitieren

Watt SA, Patschkowski T, Kalinowski J, Niehaus K. Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT). Journal of Biotechnology. 2003;106(2-3):287-300.
Watt, S. A., Patschkowski, T., Kalinowski, J., & Niehaus, K. (2003). Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT). Journal of Biotechnology, 106(2-3), 287-300. doi:10.1016/j.jbiotec.2003.07.014
Watt, S. A., Patschkowski, T., Kalinowski, J., and Niehaus, K. (2003). Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT). Journal of Biotechnology 106, 287-300.
Watt, S.A., et al., 2003. Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT). Journal of Biotechnology, 106(2-3), p 287-300.
S.A. Watt, et al., “Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT)”, Journal of Biotechnology, vol. 106, 2003, pp. 287-300.
Watt, S.A., Patschkowski, T., Kalinowski, J., Niehaus, K.: Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT). Journal of Biotechnology. 106, 287-300 (2003).
Watt, S. A., Patschkowski, Thomas, Kalinowski, Jörn, and Niehaus, Karsten. “Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT)”. Journal of Biotechnology 106.2-3 (2003): 287-300.

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