title: Determination of the dissociation constant of the NFκB p50/p65 heterodimer in living cells using fluorescence cross-correlation spectroscopy.
abstract: Fluorescence cross-correlation spectroscopy (FCCS) is a promising technique for observing and quantifying protein-protein interactions in vitro and in vivo. FCCS has emerged as a useful tool for obtaining parameters of the concentration of labeled particles, their molecular dynamics, as well as the size of their complexes. This chapter discusses aspects of preparing a biological system for FCCS experiments and suggests practical advice for performing FCCS in living cells. Moreover, we describe the method of FCCS to determine the dissociation constant of a transcription factor dimer in the living cell.
authors.affiliation: Laboratory for Nano-Bio Probes, Quantitative Biology Center, OLABB, Osaka University, Suita, Japan.
authors.forename: Manisha
authors.initials: M
authors.lastname: Tiwari
authors.forename: Masataka
authors.initials: M
authors.lastname: Kinjo
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival; D006801:Humans; D018613:Microscopy, Confocal; D052002:NF-kappa B p50 Subunit / Q000737:chemistry* / Q000235:genetics; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D013050:Spectrometry, Fluorescence / Q000379:methods*; D051996:Transcription Factor RelA / Q000737:chemistry* / Q000235:genetics; D014162:Transfection
chemical_list: D052002:NF-kappa B p50 Subunit; D051996:Transcription Factor RelA