Abstract
The SNAPf-tag is a chemical tag that allows rapid and highly specific covalent labeling of proteins even in the full complexity of the cellular environment. The SNAPf-tag has been instrumental to study native RNA-protein complexes at single-molecule resolution in their cellular environment as efficient labeling of the RNAs and proteins of interest is essential for this colocalization single-molecule spectroscopy (CoSMoS) technique. However, removal of excessive benzylguanine dye after the labeling reaction has remained challenging. Here, we describe a strategy to remove excessive benzylguanine dye using SNAPf-tag coated beads as sponges.
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Acknowledgments
Joerg E. Braun acknowledges funding from the Human Frontier Science Program (HFSP) LT000166/2013 and the European Molecular Biology Organization (EMBO) ALTF 890-2012. Joerg E. Braun is supported by funding to the laboratory of Melissa J. Moore: NIH R01 GM053007.
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Haraszti, R.A., Braun, J.E. (2020). Preparation of SNAPf-Beads for Colocalization Single-Molecule Spectroscopy (CoSMoS) of RNA-Protein Complexes. In: Arluison, V., Wien, F. (eds) RNA Spectroscopy. Methods in Molecular Biology, vol 2113. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0278-2_2
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DOI: https://doi.org/10.1007/978-1-0716-0278-2_2
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