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Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions
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Biochemistry
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JoVE Journal Biochemistry
Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions

DOI:
10.3791/60607-v

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04:45 min

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February 10, 2022

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, , , ,
1Department of Biochemistry,University of Illinois at Urbana-Champaign, 2Department of Chemistry,University of South Florida, 3Center for Biophysics and Quantitative Biology,University of Illinois at Urbana-Champaign, 4Beckman Institute for Advanced Science and Technology,University of Illinois at Urbana-Champaign

Chapters

  • 00:04Introduction
  • 00:49Microscale Thermophoresis (MST) Device Preparation
  • 01:36Labeled MST Sample Preparation
  • 02:24Sample MST
  • 02:54Results: Representative Thermophoretic Traces
  • 03:46Conclusion

Summary

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Microscale thermophoresis obtains binding constants quickly at low material cost. Either labeled or label free microscale thermophoresis is commercially available; however, label free thermophoresis is not capable of the diversity of interaction measurements that can be performed using fluorescent labels. We provide a protocol for labeled thermophoresis measurements.

Tags

Keywords: Microscale ThermophoresisProtein-lipid InteractionsVam7Phosphatidic AcidDisassociation Constant (KD)Biochemical InteractionsNTA-Atto 647 DyeThermophoretic Traces1:1 TitrationDiC8 PAPharmaceutical Development

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