É necessária uma assinatura da JoVE para visualizar este conteúdo. Faça login ou comece sua avaliação gratuita.
Capítulos
- 00:00Título
- 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
התא חדיר crosslinker DSP [dithiobis-(succinimidyl propionate)] מייצב אינטראקציות חלוף יציב<em> In vivo</em>, המאפשרת בידוד מחמירים שלהם באמצעות חלבון מורכב טכניקות לטיהור. כאן אנו מציגים טכניקה תאים crosslinking גדל בתרבות בעקבות הבידוד של קומפלקסים חלבונים על ידי 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
