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Application of Label-Free Proteomics for Differential Analysis of Lung Carcinoma Cell Line A549

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Quantitative Methods in Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 893))

Abstract

A label-free solution basing on a highly reproducible and stable LC-MS/MS system allows quantitative proteome analyses. Due to nonlabeling approach, the label-free method has the potential to measure samples from clinical specimen monitoring and comparing thousands of proteins. The presented label-free workflow includes in-solution digest, LC-MS analyses, data evaluation by the means of Progenesis™ software, and validation of the differential proteins. We successfully applied this workflow in a proteomics study analyzing the human lung carcinoma cell line A549 treated with transforming growth factor beta 1, a cell culture model of lung fibrosis. The differential analysis of only 1 μg protein per sample led to 202 significantly regulated proteins.

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References

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Acknowledgments

The author would like to thank Dr. Katja Kuhlmann and Nadine Stoepel for the mass spectrometry measurements and Kathy Pfeiffer for excellent technical assistance.

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Authors and Affiliations

  1. Department of Medical Proteomics/Bioanalytics, Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany

    Barbara Sitek, Daniel M. Waldera-Lupa & Gereon Poschmann

  2. Molecular Proteomics Laboratory, Heinrich-Heine-Universität, Düsseldorf, Germany

    Daniel M. Waldera-Lupa, Gereon Poschmann & Kai Stühler

  3. Department of Medical Proteomics/Bionalaytics, Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany

    Helmut E. Meyer

Authors
  1. Barbara Sitek
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  2. Daniel M. Waldera-Lupa
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  3. Gereon Poschmann
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  4. Helmut E. Meyer
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  5. Kai Stühler
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Corresponding author

Correspondence to Barbara Sitek .

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Editors and Affiliations

  1. Fak. Medizin, Medizinische Proteom Center, Universität Bochum, Universitätsstraße 150, Bochum, 44801, Germany

    Katrin Marcus

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© 2012 Springer Science+Business Media, LLC

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Sitek, B., Waldera-Lupa, D.M., Poschmann, G., Meyer, H.E., Stühler, K. (2012). Application of Label-Free Proteomics for Differential Analysis of Lung Carcinoma Cell Line A549. In: Marcus, K. (eds) Quantitative Methods in Proteomics. Methods in Molecular Biology, vol 893. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-885-6_16

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  • DOI: https://doi.org/10.1007/978-1-61779-885-6_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-884-9

  • Online ISBN: 978-1-61779-885-6

  • eBook Packages: Springer Protocols

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