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Ett-trinns Rensing av Twin-streptokokk-merket Proteiner og deres Komplekser for streptokokk-Tactin Resin tverrbundet Med Bis (sulfosuccinimidyl) suberat (BS3)
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Biologie
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Journal JoVE Biologie
One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)

Ett-trinns Rensing av Twin-streptokokk-merket Proteiner og deres Komplekser for streptokokk-Tactin Resin tverrbundet Med Bis (sulfosuccinimidyl) suberat (BS3)

DOI:
10.3791/51536-v

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18:27 min

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April 20, 2014

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, , ,
1Department of Food and Environmental Sciences,University of Helsinki, 2Institute of Biotechnology,University of Helsinki

Chapitres

  • 00:05Titre
  • 00:11Introduction
  • 01:32Vue d'ensemble
  • 01:37Competitive Elution with Biotin
  • 02:44Denaturing Elution with SDS
  • 03:26Chemical Cross-linking of the Resin
  • 04:10Protocol
  • 04:13Cross-linking of the Resin with BS3
  • 07:22Quenching of the Cross-linking Reaction
  • 09:50Binding of the Bait Protein and Associated Complexes to the Resin
  • 12:18Washing of the Resin
  • 14:23Elution of Specific Protein Complexes
  • 15:32Representative Results
  • 17:18Conclusion

Summary

Traduction automatique

Traduction automatique

En fremgangsmåte er beskrevet for effektiv rensing av en tvilling-Strep-merkede fusjonsproteiner og deres spesifikke komplekser på modifisert streptavidin (Strep-Tactin) harpiks kovalent kryssbundet med Bis (sulfosuccinimidyl) suberat (BS3). Fremgangsmåten har fordelen med høy hastighet, god målprotein utvinning og høy renhet, og er forenlig med etterfølgende analyse ved massespektrometri.

Tags

Keywords: Twin-Strep-tagSIII-tagStrep-TactinAffinity PurificationBis(sulfosuccinimidyl) Suberate (BS3)Cross-linkingSDS ElutionProtein ComplexesMass SpectrometryVPg-ProN. Benthamiana

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