Bioluminescence Resonance Energy Transfer (BRET)-Based Assay for Measuring Interactions of CRAF with 14-3-3 Proteins in Live Cells

873 Views

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06:44 min

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March 1st, 2024

DOI :

10.3791/66436-v

March 1st, 2024

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Russell Spencer-Smith¹^,²^,³

¹Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, ²Hollings Cancer Center, Medical University of South Carolina, ³Laboratory of Cell and Developmental Signaling, Center for Cancer Research, National Cancer Institute-Frederick

Summary

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Bioluminescence Resonance Energy Transfer BRET

14 3 3 Proteins

Ras MAPK Pathway

KRAS driven Cancers

Noonan Syndrome

Protein Interactions

Live Cell Assays

Therapeutic Approaches

Energy Transfer Protocol

Chapters in this video

0:00

Introduction

1:05

Transfection of 293FT Cells with pCMV5‐NanoLuc‐CRAF and pCMV5‐14‐3‐3ζ‐Halo Expression Constructs

3:04

Preparing 293FT Cells Transfected with pCMV5‐NanoLuc‐CRAF and pCMV5‐14‐3‐3ζ‐Halo for BRET Emission Assay

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