Testing Suitability of Cell Cultures for SILAC-Experiments Using SWATH-Mass Spectrometry
Reinders Y, Voeller D, Boßerhoff AK, Oefner PJ, Reinders J (2016)
Publication Type: Journal article
Publication year: 2016
Journal
Publisher: Humana Press, Inc.
Book Volume: 1394
Pages Range: 101-8
DOI: 10.1007/978-1-4939-3341-9_8
Abstract
Precise quantification is a major issue in contemporary proteomics. Both stable-isotope-labeling and label-free methods have been established for differential protein quantification and both approaches have different advantages and disadvantages. The present protocol uses the superior precision of label-free SWATH-mass spectrometry to test for suitability of cell lines for a SILAC-labeling approach as systematic regulations may be introduced upon incorporation of the "heavy" amino acids. The SILAC-labeled cell cultures can afterwards be used for further analyses where stable-isotope-labeling is mandatory or has substantial advantages over label-free approaches such as pulse-chase-experiments and differential protein interaction analyses based on co-immunoprecipitation. As SWATH-mass spectrometry avoids the missing-value-problem typically caused by undersampling in highly complex samples and shows superior precision for the quantification, it is better suited for the detection of systematic changes caused by the SILAC-labeling and thus, can serve as a useful tool to test cell lines for changes upon SILAC-labeling.
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Germany (DE)How to cite
APA:
Reinders, Y., Voeller, D., Boßerhoff, A.K., Oefner, P.J., & Reinders, J. (2016). Testing Suitability of Cell Cultures for SILAC-Experiments Using SWATH-Mass Spectrometry. Methods in Molecular Biology, 1394, 101-8. https://doi.org/10.1007/978-1-4939-3341-9_8
MLA:
Reinders, Yvonne, et al. "Testing Suitability of Cell Cultures for SILAC-Experiments Using SWATH-Mass Spectrometry." Methods in Molecular Biology 1394 (2016): 101-8.
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