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Chapters
- 00:05Title
- 01:37Preparation of Cell Cytosol Containing Functional Glucocorticoid Receptor
- 03:04Immunoabsorption of GR from the Cytosol and Dissociation of hsp90
- 05:16Reconstitution of GR-hsp90 Heterocomplex using Molecular Chaperones, Cofactors, and ATP
- 06:17Analysis of Reconstituted Protein Complexes and Ligand Binding Activity
- 08:26Representative GR-hsp90 Heterocomplex Formation and Steroid Binding Activity
- 10:04Conclusion
Un<em> In vitro</emMétodo> para la preparación funcional del receptor de glucocorticoides (GR) • hsp90 complejos de proteínas de las proteínas purificadas y lisados celulares se describe. El método utiliza inmunoadsorción de GR recombinante seguido por la sal-stripping y la reconstitución de proteínas complejas. La importancia de los cofactores y las condiciones de amortiguación se discuten, como lo son las posibles aplicaciones del método.
Tags
Biochemical ReconstitutionSteroid ReceptorHsp90 Protein ComplexesLigand BindingMolecular ChaperoneEukaryotic Signaling ProteinsTranscription FactorsProtein KinasesCell CyclingTumorigenesisApoptosisSignaling PathwaysGlucocorticoid Receptor (GR)Immunoprecipitation AssayIn Vitro Reconstitution MethodHsp90 Functional ActivitySteroid Receptor Ligand BindingMolecular Chaperone Cofactor Requirements
