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Chapters
- 00:00Title
- 00:41Introduction
- 01:13Preparing for Cross-linking
- 03:08Cross-linking and Cell Lysis
- 05:59Immunoprecipitation and Elution of the Bound Proteins
- 09:28IP of Cross-linked Complexes Representative Results
- 10:14Conclusion
La cellule perméables réticulant DSP [dithiobis (succinimidyl propionate)] stabilise les interactions transitoires et labiles<em> In vivo</em>, Qui permet leur isolement à l'aide des techniques rigoureuses purification des protéines recombinantes complexes. Nous présentons ici une technique pour les cellules de réticulation en culture suivie de l'isolement de complexes protéiques par immunoprécipitation.
Tags
IsolationLabile Multi-protein ComplexesIn VivoControlled Cellular Cross-linkingImmuno-magnetic Affinity ChromatographyDynamic NatureTransient Protein AssociationsWeak Protein AssociationsLow Affinity InteractionsIsolation MethodsIdentification MethodsProtein NetworksChemical CrosslinkersSelective StabilizationBiochemical LimitationsPurification MethodsProtocolCells In CultureHomobifunctional CrosslinkerSpacer ArmDithiobis-(succinimidyl Proprionate)CleavageReductionDisulphide BondCross-linking And Immunoaffinity ChromatographyMagnetic BeadsProtein ComplexesPurification TechniquesWestern Blot TechniquesMass Spectrometry
