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. Author manuscript; available in PMC: 2012 Nov 8.
Published in final edited form as: Methods Enzymol. 2012;512:223–241. doi: 10.1016/B978-0-12-391940-3.00010-X

Figure 4.1. KD determination using a fluorescence quenching approach.

Figure 4.1

Fluorescence quenching of the yAsf1*532 fluorescence signal by H3/H4Qsy9 and unlabeled H3/H4 was observed and quantitated to determine the KD of the yAsf1-H3/H4 interaction. H3/H4 or H3/H4*Qsy9 was titrated into 1.0 nM yAsf1*532. The dilution corrected and background subtracted fluorescence data were fitted with a ligand-depleted binding model (Equation 1; GraphPad Prism) because the concentration of yAsf1*532 was within 10-fold of the KD value.