A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:05Title
- 01:19Protein Purification and Alkylation of Free Sulfhydryl Groups
- 02:59Protein Digestion, Reversed Phase Peptide Extraction, and Reductive Dimethylation Labeling
- 06:02Basic pH Reversed Phase Chromatography and STop and Go Extraction
- 08:17Microcapillary LC-MS/MS and Peptide Quantification
- 09:58Results: Accuracy, Precision, and Reproducibility of ReDi Labeling
- 11:24Conclusion
Stabiele isotoop labeling van peptiden door reductieve dimethylering (Redi etikettering) is een snelle, goedkope strategie voor nauwkeurige massa-spectrometrie gebaseerde kwantitatieve proteomics. Hier laten we een robuuste werkwijze voor de bereiding en analyse van eiwitmengsels met de Redi aanpak die kan worden toegepast op vrijwel elk type monster.
Tags
Quantitative ProteomicsReductive DimethylationStable Isotope LabelingMass SpectrometryProtein Expression DifferencesRegular/formaldehyde LabelingDeuterated LabelingSodium CyanoborohydrideMethyl TagsLC-MS/MS AnalysisIon Chromatogram Peak AreasSample PreparationReversed-phase Solid Phase ExtractionBasic PH Reversed-phase ChromatographyStageTip Peptide PurificationAdvantages Of ReDi LabelingLimitations Of ReDi LabelingNovel Applications Of ReDi Labeling
