JoVE Journal

Biochemistry

You have full access to this content through

Nanyang Technological University

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

A protocol for high-precision FRET experiments at the single molecule level is presented here. Additionally, this methodology can be used to identify three conformational states in the ligand-binding domain of the N-methyl-D-aspartate (NMDA) receptor. Determining precise distances is the first step towards building structural models based on FRET experiments.

Chapters in this video

0:05

Title

1:07

Preparation of Materials

2:57

Pulsed Interleaved Excitation-Multiparameter Fluorescence Detection (PIE-MFD) Instrument Setup

6:14

Burst Integration Fluorescence Lifetime Analysis of Standards

7:34

dsDNA and Protein Sample Measurements and Analysis

9:28

Results: Inter-dye Distance Determination in Single Molecules

10:54

Conclusion

Related Videos

We use cookies to enhance your experience on our website.

By continuing to use our website or clicking “Continue”, you are agreeing to accept our cookies.

Learn More