JoVE 비디오를 활용하시려면 도서관을 통한 기관 구독이 필요합니다. 전체 비디오를 보시려면 로그인하거나 무료 트라이얼을 시작하세요.
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
Cellen genomsläppliga crosslinker DSP [dithiobis-(succinimidyl propionat)] stabiliserar övergående och labil interaktioner<em> In vivo</em>, Som gör sin isolering med hjälp av stränga proteinkomplex reningstekniker. Här presenterar vi en teknik för crosslinking celler som odlas i kultur följt av isolering av protein komplex med immunoprecipitation.
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
