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Capítulos
- 00:05Título
- 01:30Transfection
- 02:20Cell Lysis
- 03:30Binding to Rabbit IgG-agarose
- 04:53TEV Protease Cleavage
- 05:48Binding to Immobilized Streptavidin Beads
- 07:50Biotin Elution of the Streptavidin Binging Peptide and Bait Protein
Tandem affinitetsrening är en robust metod för identifiering av proteinbindning partners. Som bevis på konceptet, var denna metod tillämpas på väl karakteriserade translationsinitiering faktor eIF4E till samarbete fälla ut värdcellens faktorer inblandade i translationsinitiering. Denna metod kan lätt anpassas till någon cellulärt eller viralt protein.
Tags
Protein Interacting PartnersTandem Affinity PurificationIdentification Of Interacting ProteinsYeast Two Hybrid SystemPull Down AssaysImmunoprecipitationMass Spectrometry IdentificationDouble-affinity Tag Mediated PurificationTAPEukaryotic Translation Initiation Factor EIF4ECellular Translation Factor EIF4EEIF4F ComplexCap-dependent Translation InitiationProtein G UnitsStreptavidin Binding PeptideTobacco Etch Virus (TEV) Protease Cleavage Sequence
