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Chapters
- 00:00Title/ Prologue
- 00:56Introduction
- 01:22Separating Protein Samples by Gel Electrophoresis
- 02:06Assembling the Immunoblot Sandwich
- 05:43Transferring Proteins from Gel to Membrane
- 07:52Immunoprobing Membrane with Directly Conjugated Secondary Antibody
- 10:45Immunoprobing with Avidin-Biotin Coupling to Secondary Antibody
- 14:26Visualization of Immunoblot with Chromogenic Substrates
- 15:32Conclusion
Immunoblotting (western blotting) is a rapid and sensitive assay for the detection and characterization of proteins that works by exploiting the specificity inherent in antigen-antibody recognition. This video provides protocols for protein separation, blotting proteins onto membranes, immunoprobing, and visualization using chromogenic or chemiluminescent substrates.
Tags
Immunoblot AnalysisWestern BlottingProteinsAntigen-antibody RecognitionSolubilizationElectrophoretic SeparationGel ElectrophoresisNitrocellulosePVDFNylonSpecific AntigensPolyclonal AntibodiesMonoclonal AntibodiesSensitivityProtein SeparationMembrane BlottingImmunoprobingVisualizationChromogenic SubstratesChemiluminescent Substrates
