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A Liquid Phase Affinity Capture Assay Using Magnetic Beads to Study Protein-Protein Interaction: The Poliovirus-Nanobody Example
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JoVE Journal Biology
A Liquid Phase Affinity Capture Assay Using Magnetic Beads to Study Protein-Protein Interaction: The Poliovirus-Nanobody Example

A Liquid Phase Affinity Capture Assay Using Magnetic Beads to Study Protein-Protein Interaction: The Poliovirus-Nanobody Example

DOI:
10.3791/3937-v

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08:10 min

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May 29, 2012

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, ,
1Department of Pharmaceutical Biotechnology and Molecular Biology, Centre for Pharmaceutical Research,Vrije Universiteit Brussel

Chapters

  • 00:05Title
  • 02:27Preparation of the Magnetic Beads
  • 03:00Affinity Capture Assay
  • 04:18Recycling the Beads
  • 05:44Interpretation of the Results
  • 06:13Results: Liquid Phase Affinity Capture Assay using PVSS38C Nanobody
  • 07:36Conclusion

Summary

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In this article, a simple, quantitative, liquid phase affinity capture assay is presented. It is a reliable technique based on the interaction between magnetic beads and tagged proteins (e.g. nanobodies) on one hand and the affinity between the tagged protein and a second, labeled protein (e.g. poliovirus) on the other.

Tags

Liquid Phase Affinity Capture AssayMagnetic BeadsProtein-protein InteractionTagged ProteinLabeled ProteinCobalt Coated Magnetic BeadsRecognition Of The TagBound FractionUnbound FractionConformational Conversion Sensitive ProteinsPoliovirus-nanobody InteractionELISASolid Surface Attachment

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