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Chapters
- 00:00Title
- 00:10Introduction to Cell Surface Protein Labeling with Ettan DIGE System
- 01:12Overview of the Experiment
- 02:19Sample Preparation
- 03:13Labeling with CyDye DIGE Fluors
- 04:03Incubation and Quenching
- 04:45Cell Lysis
- 05:29Rehydration of IPG Strips
- 06:31First Dimension ISO-Electric Focusing
- 07:27Gel Casting
- 07:55Equilibration of IPG Strips
- 09:43Second Dimension SDS-Page
- 10:29Gel Scanning
- 10:52Results of Cell Surface Protein Labeling
- 11:21Comparison with Fractionated Runs
- 12:302D DIGE Analysis
- 13:27Conclusions
A simple and specific method was demonstrated for fluorescent labeling and enhanced detection of cell surface proteins without a fractionation step. Differential abundance in cell surface proteins was analyzed using two-dimensional (2-D) electrophoresis and Ettan™ DIGE technology.
Tags
Selective LabellingCell-surface ProteinsCyDye DIGE Fluor Minimal DyesSurface ProteinsIntracellular SignalingEnvironmental AdaptationDrug TreatmentDisease PathogenesisPathologyProtein-protein InteractionsSensitive MethodsReliable MethodsCell Surface Proteins Detection2-D ElectrophoresisBiomarkers DetectionComplex Protein SamplesDifferential ChangesLow Abundance ProteinsFractionationEnrichmentHydrophobic NatureHigh Molecular Weight ProteinsIntact Cells ProtocolSpecific LabelingCyDye DIGE Fluor Minimal Dyes
