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Efficient Production and Purification of Recombinant Murine Kindlin-3 from Insect Cells for Biophysical Studies
JoVE Journal
Biologia
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JoVE Journal Biologia
Efficient Production and Purification of Recombinant Murine Kindlin-3 from Insect Cells for Biophysical Studies

Efficient Production and Purification of Recombinant Murine Kindlin-3 from Insect Cells for Biophysical Studies

DOI:
10.3791/51206-v

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13:52 min

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March 19, 2014

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1Division of Structural Biology, Wellcome Trust Centre for Human Genetics,University of Oxford

Capítulos

  • 00:05Título
  • 01:13Generation of Recombinant Baculovirus
  • 04:57Amplification of Recombinant Baculovirus
  • 06:23Expression of Kindlin-3 in Baculovirus-infected Sf9
  • 07:39Purification of Recombinant Kindlin-3
  • 09:33Results: Milligram Quantitites of Recombinant Murine Kindlin-3 Produced from Insect Cells
  • 13:21Conclusion

Summary

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Kindlins are fundamental to cell adhesion through integrins but studies of them have been hampered by the difficulty encountered in expressing them recombinantly in bacterial hosts. We describe here methods for their efficient production in baculovirus-infected insect cells.

Tags

Recombinant Murine Kindlin-3Insect CellsBaculovirusSf9 CellsProtein PurificationBiophysical StudiesIntegrinsCell SignalingPlatelet AggregationLeukocyte RollingOsteoclast Formation

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