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. 2005 Feb;11(2):187–196. doi: 10.1261/rna.7950605

FIGURE 7.

FIGURE 7.

Native gel mobility assay of MSL1L2 HHRz. The RNA was renatured in 5 mM HEPES (pH 7.0) and 100 mM NaCl, fast cooled on ice, and allowed to fold in the presence of 1 mM Mg2+ prior to fractionation. (Lane 1) 2.5 μM 2′OMe substrate strand only with trace 32P-end-labeled 2′OMe substrate; (lane 2) 3 μM enzyme annealed with 2.5 μM 2′OMe substrate and trace 5′ 32P-end-labeled 2′OMe substrate; (lane 3) control lane of 1 nM 5′ end-labeled 2′OMe substrate with no enzyme strand; (lanes 4–13) a titration of 1 nM 5′ end-labeled 2′OMe substrate with 10-fold or greater cold enzyme (up to 10 μM). Two conformations for the complex are observed for all concentrations of enzyme, with an apparent Kd for complex formation of 47 nM (in 1 mM Mg2+).