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Buffer

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Submitted By gawambv
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Preparation of Buffers for Use in Enzyme Studies
(by G. Gomori)
The buffers described in this section are suitable for use either in enzymatic or histochemical studies. The accuracy of the tables is within ± 0.05 pH at 23 ºC. In most cases the pH values will not be off by more than ± 0.02 pH even at 37 ºC and at molarities slightly different from those given (usually 0.05 M).
The methods of preparation described are not necessarily identical with those of the original authors. The titration curves of the majority of the buffers recommended have been redetermined by the writer. The buffers are arranged in the order of ascending pH range.
1. Hydrochloric Acid-Potassium Chloride Buffer.
Stocks solutions
A: 0.2 M solution of KCl (14.91 g in 1 L)
B: 0.2 M HCl
50 mL of A + x mL of B, diluted to a total of 200 mL x 97.0
78.0
64.5
51.0
41.5
33.3
26.3
20.6
16.6
13.2
10.6
8.4
6.7

jun 14, 2004

pH
1.0
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
2.0
2.1
2.2

Pag.1

2. Glycine-HCl Buffer.
Stocks solutions
A: 0.2 M solution of glycine (15.01 g in 1 L)
B: 0.2 M HCl
50 mL of A + x mL of B, diluted to a total of 200 mL x 5.0
6.4
8.2
11.4

pH
3.6
3.4
3.2
3.0

x
16.8
24.2
32.4
44.0

pH
2.8
2.6
2.4
2.2

3. Phthalate-Hydrochloric Acid Buffer.
Stocks solutions
A: 0.2 M solution of potassium acid phthalate (40.48 g in 1 L)
B: 0.2 M HCl
50 mL of A + x mL of B, diluted to a total of 200 mL x 46.7
39.6
33.0
26.4
20.3

pH
2.2
2.4
2.6
2.8
3.0

x
14.7
9.9
6.0
2.63

pH
3.2
3.4
3.6
3.8

4. Aconitate Buffer.
Stocks solutions
A: 0.5 M solution of aconitic acid (87.05 g in 1 L)
B: 0.2 M NaOH
20 mL of A + x mL of B, diluted to a total of 200 mL x 15.0
21.0
28.0
36.0
44.0
52.0
60.0
68.0
76.0

jun 14, 2004

pH
2.5
2.7
2.9
3.1
3.3
3.5
3.7
3.9
4.1

x
83.0
90.0
97.0
103.0
108.0
113.0
119.0
126.0

pH
4.3
4.5
4.7
4.9
5.1
5.3
5.5
5.7

Pag.2

5. Citrate Buffer.
Stocks solutions
A: 0.1 M solution of citric acid

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