Abstract
The Mass Western describes the absolute quantification of proteins based on stable isotope labeled integral standard peptides and liquid chromatography coupled selective reaction monitoring triple quadrupole mass spectrometry (LC-SRM/MS). Here, we present a detailed workflow including tips and we discuss advantages and disadvantages of using different types of MS for absolute quantification.
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Abbreviations
- SID:
-
Stable isotope dilution
- SRM:
-
Selective reaction monitoring
- PTM:
-
Posttranslational modification
- QqQ:
-
Triple quadrupole
- LC-SRM/MS:
-
Liquid chromatography coupled selective reaction monitoring triple quadrupole mass spectrometry
- HP:
-
Heavy peptide (15N13C labeled synthetic standard)
- LP:
-
Light peptide (native peptide, no labeling)
- HCD:
-
High energy collision induced dissociation
- CE:
-
Collision energy
- LDR:
-
Linear dynamic range
- LOD:
-
Limit of detection
- LOQ:
-
Limit of quantification
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Acknowledgments
We thank Thomas Naegele for useful comments and discussions.
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Lyon, D., Weckwerth, W., Wienkoop, S. (2014). Mass Western for Absolute Quantification of Target Proteins and Considerations About the Instrument of Choice. In: Jorrin-Novo, J., Komatsu, S., Weckwerth, W., Wienkoop, S. (eds) Plant Proteomics. Methods in Molecular Biology, vol 1072. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-631-3_15
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DOI: https://doi.org/10.1007/978-1-62703-631-3_15
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