A subscription to JoVE is required to view this content. Sign in or start your free trial.
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
सेल पारगम्य डीएसपी crosslinker [dithiobis (succinimidyl propionate)] स्थिर क्षणिक और अस्थिर बातचीत<em> Vivo में</em>, जो उनके अलगाव की अनुमति देता कड़े प्रोटीन जटिल शोधन तकनीक का उपयोग. यहाँ हम crosslinking संस्कृति में विकसित कोशिकाओं immunoprecipitation द्वारा प्रोटीन परिसरों के अलगाव के बाद के लिए तकनीक मौजूद है.
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
