This title appears in the Scientific Report :
2018
Please use the identifier:
http://dx.doi.org/10.21769/BioProtoc.2881 in citations.
Preparation of Cell-free Synthesized Proteins Selectively Double Labeled for Single-molecule FRET Studies
Preparation of Cell-free Synthesized Proteins Selectively Double Labeled for Single-molecule FRET Studies
Single-molecule FRET (smFRET) is a powerful tool to investigate molecular structures and conformational changes of biological molecules. The technique requires protein samples that are site-specifically equipped with a pair of donor and acceptor fluorophores. Here, we present a detailed protocol for...
Saved in:
Personal Name(s): | Sadoine, Mayuri |
---|---|
Cerminara, Michele / Fitter, Jörg (Corresponding author) / Katranidis, Alexandros (Corresponding author) | |
Contributing Institute: |
Molekulare Biophysik; ICS-5 |
Published in: | Bio-protocol, 8 (2018) 12, S. 1 |
Imprint: |
Sunnyvale, CA
bio-protocol.org
2018
|
DOI: |
10.21769/BioProtoc.2881 |
Document Type: |
Journal Article |
Research Program: |
Functional Macromolecules and Complexes |
Publikationsportal JuSER |
Single-molecule FRET (smFRET) is a powerful tool to investigate molecular structures and conformational changes of biological molecules. The technique requires protein samples that are site-specifically equipped with a pair of donor and acceptor fluorophores. Here, we present a detailed protocol for preparing double-labeled proteins for smFRET studies. The protocol describes two cell-free approaches to achieve a selective label scheme that allows the highest possible accuracy in inter‐dye distance determination. |