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High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
Chapters
- 00:05Title
- 01:07Preparation of Materials
- 02:57Pulsed Interleaved Excitation-Multiparameter Fluorescence Detection (PIE-MFD) Instrument Setup
- 06:14Burst Integration Fluorescence Lifetime Analysis of Standards
- 07:34dsDNA and Protein Sample Measurements and Analysis
- 09:28Results: Inter-dye Distance Determination in Single Molecules
- 10:54Conclusion
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.
