Rivista
/
Biochimica
/
Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription
JoVE Journal
Biochimica
È necessario avere un abbonamento a JoVE per visualizzare questo.  Accedi o inizia la tua prova gratuita.
JoVE Journal Biochimica
Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

DOI:
10.3791/59687-v

•

07:18 min

•

July 26, 2019

•

, ,
1Department of Pharmacology, Robert Wood Johnson Medical School,Rutgers University, 2Graduate Program in Physiology and Integrative Biology, School of Graduate Studies,Rutgers University, 3Department of Plant Biology, School of Environmental and Biological,Rutgers University

Capitoli

  • 00:04Titolo
  • 00:59Biosensor Hydration and Assay Set-Up
  • 02:51Loading of Ligand onto Biosensor and Washing Away Additional Ligand
  • 03:43Association of Analyte to Ligand and Dissociation of Analyte from Ligand
  • 05:41Results: A 28 Amino Acid Middle Region of GrgA Binds σ28 In Vitro
  • 06:45Conclusion

Summary

Traduzione automatica

Traduzione automatica

Interactions of transcription factors (TFs) with the RNA polymerase are usually studied using pulldown assays. We apply a Biolayer Interferometry (BLI) technology to characterize the interaction of GrgA with the chlamydial RNA polymerase. Compared to pulldown assays, BLI detects real-time association and dissociation, offers higher sensitivity, and is highly quantitative.

Tags

Biolayer InterferometryBLIProtein-protein InteractionsTranscriptionKinetic ParametersBiosensorNickel NTAReal-time Association And DissociationQuantitative AnalysisPor Dialysis

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics

Preparation of Keratin Hydrolysate from Chicken Feathers and Its Application in Cosmetics

Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Evaluation of Protein–Protein Interactions using an On-Membrane Digestion Technique

Interactions with and Membrane Permeabilization of Brain Mitochondria by Amyloid Fibrils

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions

Read Article