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GST-Hans oprensning: En to-trins Affinitetsrensning protokollen Efterlevelse fuld længde oprensede proteiner
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GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins

GST-Hans oprensning: En to-trins Affinitetsrensning protokollen Efterlevelse fuld længde oprensede proteiner

DOI:
10.3791/50320-v

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11:22 min

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October 29, 2013

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, , , , ,
1Genome Stability Laboratory, Oncology Axis,Hôtel-Dieu de Québec

Chapters

  • 00:05Title
  • 01:49Recombinant Protein Expression and Preparation of Soluble Cell Lysate
  • 03:17GST Affinity Purification
  • 05:14TALON Metal Affinity Purification
  • 07:00Results: Analysis of GST-His Purification
  • 10:21Conclusion

Summary

Automatic Translation

Automatic Translation

I den foreliggende protokol, viser vi en yderst effektiv og omkostningseffektiv små proteinoprensning metode, som tillader oprensning af rekombinante proteiner ved entydigt at kombinere en spaltelig GST-tag og en lille His-mærke.

Tags

GST-His PurificationTwo-step Affinity Purification ProtocolFull-length Purified ProteinsProtein PurificationHigh ConcentrationsBiochemical CharacterizationEnzymology AssaysEfficiencySimplicityCost EffectivenessN-terminal Glutathione Sepharose Tag (GST)C-terminal 10xHis TagRecombinant ProteinsBaculovirusesSf9 Infected CellsProtein ExpressionPreScission EnzymeMaximum PurityCleaved GST RemovalHis-Tag Affinity PurificationDegraded Proteins RemovalFull-length Proteins EnrichmentInstrumental SetupFPLCMgCl2 WashesATP WashesHeat Shock Protein Impurities Removal

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