You need a buff er at pH 7.5 for use in purifying a protein at 4°C. You have chosen Tris, pK 8.08, ΔH° = 50 kJ · mol−1. You carefully make up 0.01 M Tris buffer, pH 7.5 at 25°C, and store it in the cold to equilibrate it to the temperature of the purifi cation. When you measure the pH of the temperature-equilibrated buff er it has increased to 8.1. What is the explanation for this increase? How can you avoid this problem?

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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You need a buff er at pH 7.5 for use in purifying a protein at 4°C. You have chosen Tris, pK 8.08, ΔH° = 50 kJ · mol−1. You carefully make up 0.01 M Tris buffer, pH 7.5 at 25°C, and store it in the cold to equilibrate it to the temperature of the purifi cation. When you measure the pH of the temperature-equilibrated buff er it has increased to 8.1. What is the explanation for this increase? How can you avoid this problem?

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