Biology Reference
In-Depth Information
are used, SDS concentration cannot exceed 0.1 % SDS.
Consequently, samples must be diluted with HENU before
loading on the fi lter devices. Urea in HENU buffer is used to
facilitate SDS removal.
9. A control of the labeling specifi city is absolutely necessary.
Indeed, potential incomplete free thiol blocking needs to be
evaluated. It is then recommended to introduce an additional
control where the nitrosothiol reduction by ascorbate/CuCl
2
is omitted.
10. Injecting the Elute buffer before loading sample is required to
free up low-affi nity binding sites on the avidin cartridge.
11. If TP is true positive matches and FP is false positive matches,
the number of matches in the target database is TP + FP and
the number of matches in the decoy database is FP. The quan-
tity that is reported is the False Discovery Rate (FDR) = FP/
(FP + TP).
Acknowledgments
A.F. benefi ted from a PhD grant of the Région Languedoc-
Roussillon. The authors acknowledge the help of the INRA Mass
Spectrometry Proteomics Platform from the Pole Protéome de
Montpellier in performing the analysis.
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