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. 2010 Dec 31;285(53):le26–le28. doi: 10.1074/jbc.N110.160093

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Isothermal titration calorimetry investigation of the influence of glycerol in various buffers in buffer to buffer experiments. ITC was conducted by injecting a specific buffer/glycerol formulation into a sample well containing the same buffer/glycerol formulation. Buffers were: 10 mm phosphate, 100 mm NaCl, 50 mm AMS, pH 6.5 (A); 10 mm phosphate, 100 mm NaCl, 50 mm AMS, 2.5% glycerol (v/v), pH 6.5 (B); 10 mm phosphate, 100 mm NaCl, 50 mm AMS, 5% glycerol (v/v), pH 6.5 (C); ITC Buffer (Falk et al. (3)): 20 mm HEPES/NaOH pH 7.5, 200 mm NaCl, 2 mm MgCl₂, 1 mm EDTA, 0.25 mm TCEP (D); ITC Buffer with 2.5% glycerol (Falk et al. (3)): 20 mm HEPES/NaOH pH 7.5, 200 mm NaCl, 2 mm MgCl₂, 1 mm EDTA, 0.25 mm TCEP, 2.5% glycerol (v/v) (E); and ITC Buffer with 5% glycerol (Falk et al. (3)): 20 mm HEPES/NaOH pH 7.5, 200 mm NaCl, 2 mm MgCl₂, 1 mm EDTA, 0.25 mm TCEP, 5% glycerol (v/v) (F). Although polyols such as glycerol are frequently used to stabilize proteins, they cannot be assumed innocuous.