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Technical, Experimental, and Biological Variations in Isobaric Tags for Relative and Absolute Quantitation (iTRAQ)
Abstract
We assess the reliability of isobaric-tags for relative and absolute quantitation (iTRAQ), based on different types of replicate analyses taking into account technical, experimental, and biological variations. In total, 10 iTRAQ experiments were analyzed across three domains of life involving Saccharomyces cerevisiae KAY446, Sulfolobus solfataricus P2, and Synechocystis sp. PCC 6803. The coverage of protein expression of iTRAQ analysis increases as the variation tolerance increases. In brief, a cutoff point at +/-50% variation (+/-0.50) would yield 88% coverage in quantification based on an analysis of biological replicates. Technical replicate analysis produces a higher coverage level of 95% at a lower cutoff point of +/-30% variation. Experimental or iTRAQ variations exhibit similar behavior as biological variations, which suggest that most of the measurable deviations come from biological variations. These findings underline the importance of replicate analysis as a validation tool and benchmarking technique in protein expression analysis.
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Cited by
Park, J. E., Tan, H. S., Datta, A., Lai, R. C., Zhang, H., Meng, W., Lim, S. K., & Sze, S. K. (2010) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m900381-mcp200 · Cited by 582
iTRAQ Underestimation in Simple and Complex Mixtures: “The Good, the Bad and the Ugly”
Ow, S. Y., Salim, M., Noirel, J., Evans, C., Rehman, I., & Wright, P. C. (2009) · Journal of Proteome Research · doi:10.1021/pr900634c · Cited by 489
Self-citationA cytosolic network suppressing mitochondria-mediated proteostatic stress and cell death
Wang, X., & Chen, X. J. (2015) · Nature · doi:10.1038/nature14859 · Cited by 374
Detecting significant changes in protein abundance
Kammers, K., Cole, R. N., Tiengwe, C., & Ruczinski, I. (2015) · EuPA Open Proteomics · doi:10.1016/j.euprot.2015.02.002 · Cited by 311
Current trends in quantitative proteomics
Elliott, M. H., Smith, D. S., Parker, C. E., & Borchers, C. (2009) · Journal of Mass Spectrometry · doi:10.1002/jms.1692 · Cited by 271
Metadata
- Authors
- Gan, C. S., Chong, P. K., Pham, T. K., & Wright, P. C.
- Venue
- Journal of Proteome Research
- Publisher
- American Chemical Society (ACS)
- Published
- 2007-01-05
- Volume
- 6
- Issue
- 2
- Pages
- 821-827
- ISSN
- 1535-3893 (ISSN-L)
- 1535-3907
- OpenAlex
- W2063001525
- Cited by
- 404
- References
- 0
- FWCI
- 12.49
- Type
- journal article
- Language
- en
- Open access
- closed
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- Gan, C. S., Chong, P. K., Pham, T. K., & Wright, P. C. (2007). Technical, Experimental, and Biological Variations in Isobaric Tags for Relative and Absolute Quantitation (iTRAQ). Journal of Proteome Research. https://doi.org/10.1021/pr060474i
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