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Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
Chapters
- 00:05Title
- 01:14Protein Extraction and Generation of Peptides for Chemical-tagging
- 02:05Sample Digestion
- 03:16Dimethylation Labeling (N-termini)
- 05:13Isobaric Tagging (Lys residues)
- 06:12Liquid Chromatography – Tandem Mass Spectrometry (LC-MS/MS) and MS3
- 07:02Results: cPILOT Can Be Used to Analyze Tissues from an Alzheimer’s Disease Mouse Model and Wild-type Controls
- 08:23Conclusion
Combined precursor isotopic labeling and isobaric tagging (cPILOT) is a quantitative proteomics strategy that enhances sample multiplexing capabilities of isobaric tags. This protocol describes the application of cPILOT to tissues from an Alzheimer's disease mouse model and wild-type controls.
Tags
Enhanced Sample MultiplexingCombined Precursor Isotopic LabelingIsobaric TaggingCPILOTAlzheimer’s DiseaseMouse ModelProtein ChangesAgingNeurodegenerative DiseaseCancerPathogenesisDiagnosisPrognosisTherapeutic TargetsAcetylationProtein NitrationBrainHeartLiverBCA AssayProtein ConcentrationDithiothreitolIodoacetamideL-cysteineTrypsin Digestion
