SpringerProtocols: Abstract: Fluorescence Correlation Spectroscopy (FCS)-Based Characterisation of Aptamer Ligand Interaction

7. Fluorescence Correlation Spectroscopy (FCS)-Based Characterisation of Aptamer Ligand Interaction

By: Arne Werner², Ulrich Hahn²

Abstract

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Fluorescence correlation spectroscopy (Bacia and Schwille (2007) Nat. Protoc. 2, 2842–2856) reveals molecular mobilities, enabling to identify molecular interactions based on a change of diffusion times (Rigler and Elson, (2001) Fluorescence Correlation Spectroscopy: Theory and Applications. Springer, Berlin; Haustein, and Schwille, (2004) Curr. Opin. Struct. Biol. 14, 531–540). This technique can be applied to determine the dissociation constant of a complex formed by a fluorescence-labelled target and its corresponding RNA aptamer selected via systematic evolution of ligands by exponential enrichment (SELEX) (Schürer, et al. (2001) Biol. Chem. 382, 47948). Here, an FCS titration experiment is described in detail, where the binding properties of tetramethylrhodamine (TMR) labelled Moenomycin A to its corresponding RNA aptamer were determined (Schürer, et al. (2001) Biol. Chem. 382, 47948).

Affiliation(s): (2) Department of Chemistry, Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany

Book Title: Nucleic Acid and Peptide Aptamers: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 535 | Pub. Date: Mar-03-2009 | Page Range: 1-8 | DOI: 10.1007/978-1-59745-557-2_7

Subject: Cell Biology

Key Words: Fluorescence correlation spectroscopy (FCS) - RNA aptamer, Moenomycin A - tetramethylrhodamine (TMR)

References

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