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GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins
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Biologia
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JoVE Journal Biologia
GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins

GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins

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

Capítulos

  • 00:05Título
  • 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

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In the present protocol, we demonstrate a highly efficient and cost-effective small-scale protein purification method, which allows purification of recombinant proteins by uniquely combining a cleavable GST-tag and a small His-tag.

Tags

GST-His PurificationAffinity PurificationProtein Purification ProtocolRecombinant ProteinsGlutathione Sepharose Tag (GST)10xHis TagBaculovirus ExpressionSf9 CellsPreScission EnzymeFull-length ProteinsHeat Shock Protein ImpuritiesNuclease Treatment

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