BUFFERS

BUFFERS

Ph.D.

pH pK a

pK a

pK a

11

12

pH

14

pK a pK a /

15

pH

15

16

: pH

16

pH

17

18

20

23

25

27

pH

27

pK a

28

28

29

30

32

pH pH

pH
pH

Calbiochem


70%

H OH
H OH

1Na Cl


H OH

K eq

25 55.5 M 1000 18; 18.0

K eq 25 1.8 10-16 M

[H ][OH ]25 1.0 10-14M2 [H ]

[OH ] pH

H OH
pH

k 1 k 2
-d[CH3COOH]/dt
k 1 [CH3COOH]

d[CH3COOH]/dt k 2

pH pK a
pHpK a
[H+] 10-1 M 10-10 M
S.P.L. Sorenson
"p" [H+]

pK a -logK a -log

 pH pK a

log[CH 3 COO - ]/[CH 3 COOH]

pK a 4.745
1 pH pK
a
pH 4.75
pK a pH
pK pK a pK a a
acid pH
pK b b base
pOH

pK a
pK a

pH 2 pK a

pK a
2 pK a 3
5
3 pK a1pK a2 pK a3 2 H2PO4- HPO42

 1 pK a

115226
115227
391334
391336
391335
394111
203667
205541
239782
239779
231211
3570
357002
3630
391340
391338
375368
391333
115230
4015
475893
475894
475898
475899
110220
528131
528132
528315
529567
529568
567550
567547
567545
567549
394675
39465
110695
39468
648310
648311
648317
648313
567446
1.
2.
3.

ACES
ADA
BESULTROL Grade
BicineULTROL Grade
BIS-TrisULTROL Grade
BIS-Tris ULTROL Grade

CAPSULTROL Grade
CHESULTROL Grade

HEPES
HEPESULTROL Grade
HEPESULTROL Grade
HEPESULTROL Grade
HEPPSULTROL Grade
ULTROL Grade
MESULTROL Grade
MESULTROL Grade
MOPSULTROLGrade
MOPSULTROL Grade
PIPES
PIPESULTROL Grade
PIPESULTROL Grade
PIPPS

TAPSULTROL Grade
TESULTROL Grade
TESULTROL Grade
ULTROL Grade

ULTROL

ULTROL

pK a20

182.2
212.2
213.2
163.2
209.2
282.4
61.8
214
221.3
207.3
210.1
75.1
75.1
132.1
238.3
238.3
238.3
260.3
252.3
68.1
195.2
217.2
209.3
231.2
302.4
302.4
335.8
330.4
228.2
136.1
142
142
120
138
243.2
229.3
251.2
179.2
121.1
121.1
157.6
157.6
294.1

6.9
6.6
7.15
8.35
6.5
6.8
9.24
6.27
10.4
9.5
4.76
2.341
2.341
8.4
7.55
7.55
7.55
7.55
8
7
6.15
6.15
7.2
7.2
6.8
6.8
6.8
3.732
7.213
7.213
7.213
7.213
7.213
7.213
8.4
7.5
7.5
8.15
8.3
8.3
8.3
8.3

pK a1 = 2.34; pK a2 = 9.60

pK a1 = 3.73; pKa2 = 7.96(25100mM
pH
pK a pK a1 = 2.12pK a2 = 7.21pK a3 = 12.32

pH

pH pH
pH pK a

pH
1L 1 mol pH

pK a -

pK a pK a 1.0 pH

pH

pH
pH = pK a
HApK a = 7.0 [A-] [H ]

pH 7.0 10 mM1 10-2 MHCl 100 mL 1.0 M NaCl 1.0

1.1 [H+]


Vol0 = HCl
[H+]0 = M
Vol = HCl NaCl
[H+] = HCl NaCl M
[H+] :

1.0 10-3 M 4 pH 7.0

3.04
pH 7 1.0 M HA
:

1.0 10-2 M10 mMHCl 100 mL 1.0 10-3 M A- 1.0

10-3 M HA :

4 pH
pH pH
[A- ]/[HA] = 0.1 [A- ]/[HA] = 10.0

10

pK a 6.0 8.0

Good
pH pK a

11

pK a 6.86 pH 6.4 7.4

H2CO3
pH

pH 7.4
4
pH
pCO2 pH

12

 pCO2
pH

H+Cl- Na+

13

pH
2
a 1
b K a
pH

5 pH

pK a
pK a/ 2
pK a pK a/

5 pH

14

 2 pK a pK a/

pK a(20 )

pK a/

MES
ADA

195.2
212.2

6.15
6.60

-0.011
-0.011

BIS-Tris
PIPES
ACES

282.4
342.3
182.2

6.80
6.80
6.90

-0.016
-0.009
-0.020

BES

213.3

7.15

-0.016

MOPS
TES

209.3
229.3

7.20
7.50

-0.006
-0.020

HEPES
HEPPS

238.3
252.3
179.2

7.55
8.00
8.15

-0.014
-0.007
-0.021

Bicine

121.1
163.2

8.30
8.35

-0.031
-0.018

CHES
CAPS

132.1
207.3
221.3

8.40
9.50
10.40

-0.028
-0.009
-0.009

Cu2+Ca2+Mn2+ Mg2+

Cu2+ Ca2+Mn2+Mg2+

Cu2+ Ca2+Mn2+Mg2+

Cu2+ Ca2+Mn2+
Mg2+

Cu2+ Ca2+Mn2+Mg2+

Cu2+ Ca2+Mn2+Mg2+

Cu2+ Mn2+

pH

ACES
BESTES
HEPESHEPPS
Folin
Bradford

15


pH

25 1.0 10-14
1.45 10-17
pH DMSO
pH

pH
pH
pH
:

Mg(OH)2
OH- Mg(OH)2
OH- Mg(OH)2

16

pH
1.

pH

2.

pH

3.
4.
5. 2 pH
6.

7.
8.
9.

pH

10.

11.

12.

17


1. pK a pK a pH

pK a pH

pH pK a

pH pH pK a

pK a

2. pH pK a pH

pH
3.

4.

5.

240nm 700nm

6.

7.

pH 10:1 1:0

0.1 M HEPES 0.1M HEPES

pH 6.55 pH 8.55 0.1M HEPES
8.

1

pH 20 24

18

9. pH

pH pKa

pH pH pKa

pH

pH
10.

11.

6 pH [A- ]/[HA]

pH
pH
20 pH 7.6 0.1M MOPS MOPS
pK a 20 7.2 pH 0.4 pH
pK a pH MOPS /MOPS 2.5
0.1M MOPS 0.1M MOPS
pH pH

pH

6 pH [A- ]/[HA]

19


pH
pH pH
pH

1. - HCl-KCl; pH 1.0 2.2

a0.2 M : 14.9 g/L 74.5
b0.2 M
50mL
200 mL pH pH
mL

97.0

64.5

41.5

26.3

16.6

10.6

6.7

pH

1.0

1.2

1.4

1.6

1.8

2.0

2.2

2. - ; pH 2.2 3.6
a0.1 M : 7.5 g/L 75.0
b0.1 M
50 mL 100 mL
pH pH
mL

44.0

32.4

24.2

16.8

11.4

8.2

6.4

5.0

pH

2.2

2.4

2.6

2.8

3.0

3.2

3.4

3.6

3. ; pH 3.0 6.2
a0.1 M : 19.21 g/L 192.1
b0.1 M : 29.4 g/L 294.0
100 mL
pH pH

mL

46.5

40.0

35.0

31.5

25.5

20.5

16.0

11.8

7.2

mL 3.5

10.0

15.0

18.5

24.5

29.5

34.0

38.2

42.8

pH

3.4

3.8

4.2

4.6

5.0

5.4

5.8

6.2

20

3.0

4. ; pH 3.6 5.6
a0.1 M 5.8 mL 1000 mL
b0.1 M : 8.2 g/L 82.0 13.6 g/L

136.0

100 mL
pH pH
mL

46.3

41.0

30.5

20.0

14.8

10.5

4.8

mL

3.7

9.0

19.5

30.0

35.2

39.5

45.2

pH

3.6

4.0

4.4

4.8

5.0

5.2

5.6

5. - ; pH 2.6 7.0
a0.1 M : 19.21 g/L 192.1
b0.2 M : 35.6 g/L 178.0

53.6 g/L ; 268.0

100 mL pH pH
mL

44.6 39.8 35.9 32.3 29.4 26.7 24.3 22.2 19.7 16.9 13.6

6.5

mL 5.4

10.2 14.1 17.7 20.6 23.3 25.7 27.8 30.3 33.1 36.4 43.6

pH

3.0

2.6

3.4

3.8

4.2

4.6

5.0

5.4

5.8

6.2

6.6

7.0

6. ; pH 5.8 8.0
a0.2 M : 27.6 g/L 138.0
b0.2 M : 53.6 g/L 268.0

200 mL pH pH

mL 92.0

81.5

73.5

62.5

51.0

39.0

28.0

19.0

13.0

8.5

5.3

mL 8.0

18.5

26.5

37.5

49.0

61.0

72.0

81.0

87.0

91.5

94.7

pH

6.2

6.4

6.6

6.8

7.0

7.2

7.4

7.6

7.8

8.0

5.8

21

7. - ; pH 7.2 9.0
a0.1 M : 12.1 g/L 121.0
b0.1 M
50 mL
100 mL pH pH

mL

44.2

41.4

38.4

32.5

21.9

12.2

5.0

pH

7.2

7.4

7.6

7.8

8.2

8.6

9.0

8. - ; pH 8.6 10.6
a0.1 M : 7.5 g/L 75.0
b0.1 M : 4.0 g/L 40.0
50 mL
100 mL pH pH
mL

4.0

8.8

16.8

27.2

32.0

38.6

45.5

pH

8.6

9.0

9.4

9.8

10.0

10.4

10.6

9. - ; pH 9.2 10.6
a0.1 M : 10.6 g/L 106.0
b0.1 M : 8.4 g/L 84.0

100 mL pH pH

mL

4.0

mL 46.0

pH

22

9.2

9.5

16.0

22.0

27.5

33.0

38.5

42.5

40.5

34.0

28.0

22.5

17.0

11.5

7.5

9.4

9.6

9.8

10.0

10.2

10.4

10.6


1. Krebs-Henseleit , pH 7.4
a119 mM NaCl

d4.7 mM KCl

b2.5 mM CaCl2

e1.2 mM MgSO4

c1.2 mM KH2PO4

f 25 mM NaHCO3

95% O2 5% CO2
pH 7.4 (at 37 ) pH

2. Hank's , pH 7.4
a137 mM NaCl

e5.4 mM KCl

b0.25 mM Na2HPO4

f 0.44 mM KH2PO4

c1.3 mM CaCl2

g1.0 mM MgSO4

d4.2 mM NaHCO3
95% O2 5% CO2 pH 7.4 (at 37 )

3. PBS , pH 7.4
a150 mM NaCl

b10 mM

(PBS 1L 1L NaCl 8.7 g, K2HPO4.3H2O 1.82 g, KH2PO4 0.23 g

pH )
PBS :
a137 mM NaCl

c2.7 mM KCl

b10 mM Na2HPO4

d1.76 mM KH2PO4

4. TBS, pH 7.4
a10 mM Tris

b150 mM NaCl

(TBS 1L 1L Tris base 1.21 gNaCl 8.7 g pH

Tris pK a 8.3 pH 7.4
(pK a= 7.21)

23

5. TBST (TWEEN-20 )
a10 mM Tris-HCl, pH 8.0

c0.1% TWEEN 20

b150 mM NaCl

6.
a150 mM NaCl 62.5 mM Tris buffer, pH 6.7 6.8
b150 mM NaCl 100 mM -
c150 mM NaCl 2% (SDS)

7.
a20 mM Tris-HCl (pH 7.5)

f 150 mM NaCl

b1 mM Sodium EDTA

g1 mM EGTA

c1% TRITON X-100

h2.5 mM

d1 mM -

i 1 mM

e1 g/mL

24

4.5

1.8

Pencillium duponti

3.9

4.7-5.1

5.9-6.1

7.5-8.0

8.5-9.0

4.6-5.3

8.7-9.3

4.4

3.9

9.4

9.2

ATP Na+-K

5.1

6.9

6.0

6.7

5.1

6.7

7.8

8.8

Clostridium

5.5

cAMP-

6.1

7.4

DNA

4.7

DNA

4.7

4.9

5.9

4.8-5.0

5.0-5.6

4.8

4.6

-16 -

5.9

5.8

6.3

- 6 -

8.5

8.5
8.2-8.6

8.4
6.2-6.5

4.6

5.8-7.0
6.7

25

S -

6.9-8.1

3.0
8.3

D - 3 -

L - - 3 -

6.4

6.3

7.3

5.1

7.0

7.0

5.3

5.7

8.5

4.5

4.7

5.1

4.1

6.1

5.0

3 -

6.4

10.5

C. perfringens

5.3

5.8

2.2

7.0-8.5

6.4-8.5

8.9

6.5

7.5

5.3

9.3

RNA II

HeLaKB

4.8

Pleurotus olearius

7.0

7.1

5.9

4.5

8.7

5.5

4.9

26

5.4
6.8-7.3

7.8
4.6-4.7
5.7

66,500

5.2

44,000

2.7

51,000

4.2-4.7

135,000

4.4

320,000

5.9

110,000

4.8

4.8-5.0
4.8

70

340,000

5.5

IgG

150,000

5.8-7.3

IgD

172,000

4.7-6.1

44

5.2

725,000

5.4

11,800

5.8

7.0-8.5

50.000-60,000

4.7

4.6-4.9

37,000

7.1

63,000

4.2-5.2

79,600

5.9

5 pH

pH

meqHCO3-/l

7.35-7.45

28

7.4

25

6.4-7.4

10-20

1.0-2.0

7.0-7.4

5-25

7.4

28

7.0-8.0

80

4.5-7.5

0-10

27

 6 pK a

pK a

1.75 10-5

4.76

7.4 10-4
1.7 10-5
4.0 10-7

3.13
4.77
6.40

1.75 10-4

3.75

4.5 10-3
1.7 10-10

2.35
9.77

1.01 10-7

6.95

7.5 10-3
6.2 10-8
4.8 10-13

2.12
7.21
12.32

3.23 10-3

2.49

8.32 10-9

8.08

1N1LmL

60.05

1.06

99.5

17.6

57

36.46

1.19

37

12.1

83

36.02

1.42

70

15.7

64

72

100.46

1.68

72

11.9

84

98.00

1.70

85

44.1

23

98.08

1.84

96

36.0

28

28

A = abc

A =

a =

b = cm

c =
b 1 cm c mol/La
pH

:
25 60,700
584
1 cm 5 mg/L0.005 g/L
A =60,70010.005/584= 0.52 {A = abc}
0.49 94%0.49/0.52
mol/L g/dl

1%
b = 1 cm; c = 1 g/dl1%
A A
1cm

K
A = abc

c =

A A 1
ab
ab

ab 1/ab K

c = A K; K = c/A
K

29

 Calbiochem
ULTROL

ULTROL

115226

ACES

115227

ADA, Monosodium Salt

391334

BES, Free Acid, ULTROL Grade

391336
394111
391335

Bicine, ULTROL Grade

BIS-Tris Propane, ULTROL Grade
BIS-Tris, ULTROL Grade

203667

Boric Acid, Molecular Biology Grade

205541
239782

Cacodylic Acid, Sodium Salt

CAPS, ULTROL Grade

239779
231211
3570

CHES, ULTROL Grade

Citric Acid, Monohydrate, Molecular Biology Grade
Glycine

357002

Glycine, Molecular Biology Grade

115226-100GM
115226-250GM
115226-10KG
115227-100GM
115227-250GM
115227-10KG
391334-25GM
391334-100GM
391336-100GM
394111-100GM
391335-100GM
391335-1KG
203667-500GM
203667-1KG
203667-5KG
205541-100GM
239782-100GM
239782-1KG
239779-100GM
231211-100GM
3570-500GM
3570-1KG
357002-100GM
357002-1KG
3630-25GM
3630-100GM
391340-25GM
391340-250GM
391338-25GM
391338-100GM
391338-500GM
391338-1KG
391338-5KG
375368-100ML
375368-500ML
391333-100GM
391333-500GM
391333-1KG
115230-25GM
115230-250GM
115230-10KG
4015-25GM
4015-100GM
475893-100GM
475893-500GM
475893-1KG
475894-100GM
475894-1KG
475898-100GM
475898-500GM
475898-1KG

3630
391340

HEPES, Free Acid, Molecular Biology Grade

391338

HEPES, Free Acid, ULTROL Grade

375368

HEPES, Free Acid, ULTROL Grade 1M-Solution

391333

HEPES, Sodium Salt, ULTROL Grade

115230

HEPPS, ULTROL Grade

4015

30

Glycylglycine, Free Base

Imidazole, ULTROL Grade

475893

MES, Free Acid, ULTROL Grade

475894

MES, Sodium Salt, ULTROL Grade

475898

MOPS, Free Acid, ULTROL Grade

100 g
250 g
10 Kg
100 g
250 g
10 kg
25 g
100 g
100 g
100 g
100 g
1k g
500 g
1 kg
5 kg
100 g
100 g
1 kg
100 g
100 g
500 g
1 kg
100 g
1 kg
25 g
100 g
25 g
250 g
25 g
100 g
500 g
1k g
5kg
100 mL
500 mL
100 g
500 g
1 kg
25 g
250 g
10 kg
25 g
100 g
100 g
500 g
1 kg
100 g
1 kg
100 g
500 g
1 kg

475899

MOPS, Sodium Salt, ULTROL Grade

475916

MOPS/EDTA Buffer, 10X Liquid Concentrate, Molecular Biology

Grade
PBS Tablets
PBS-TWEENTablets
PIPES, Free Acid, Molecular Biology Grade

475899-100GM
475899-1KG
475916-100ML

524650
524653
110220

528131
528132
528315
529567
529568
567446

567550
567547
567545
567549
567780
567784
106023
394675
574796
574800
39465
110695

585205
585207
39468
625718
648310
648314
648315
648317
648313

648311

524650-1EA
524653-1EA
110220-25GM
110220-250GM
110220-1KG
110220-10KG
PIPES, Free Acid, ULTROL Grade
528131-100GM
PIPES, Sesquisodium Salt, ULTROL Grade
528132-100GM
528132-1KG
PIPPS
528315-100GM
Potassium Phosphate, Dibasic, Trihydrate, Molecular Biology Grade 529567-250GM
529567-1KG
Potassium Phosphate, Monobasic, Molecular Biology Grade
529568-250GM
529568-1KG
Sodium Citrate, Dihydrate, Molecular Biology Grade
567446-100GM
567446-1KG
567446-5KG
Sodium Phosphate, Dibasic
567550-500GM
567550-1KG
Sodium Phosphate, Dibasic, Molecular Biology Grade
567547-250GM
567547-1KG
Sodium Phosphate, Monobasic
567545-500GM
567545-1KG
Sodium Phosphate, Monobasic, Monohydrate, Molecular Biology
567549-250GM
Grade
567549-1KG
SSC Buffer, 20X Powder Pack, ULTROL Grade
567780-2PACK
SSPE Buffer, 20X Powder Pack, ULTROL Grade
567784-2PACK
TAE Buffer, 10x pH 8.3 tris-acetate-EDTA buffer
106023-1L
TAPS, ULTROL Grade
394675-1KG
TBE Buffer, 10X Powder Pack, ULTROL Grade
574796-2PACK
TE Buffer, pH 8.0, Molecular Biology Grade
574800-100ML
TES, Free Acid, ULTROL Grade
39465-100GM
39465-1KG
TES
110695-25GM
110695-250GM
110695-10KG
TG Buffer, 10X Powder Pack, ULTROL Grade
585205-2PACK
TG-SDS Buffer, 10X Powder Pack, ULTROL Grade
585207-2PACK
Tricine, ULTROL Grade
39468-100GM
39468-1KG
Triethylammonium Acetate, 1M Solution
625718-1L
Tris Base, Molecular Biology Grade
648310-500GM
648310-2.5KG
Tris Buffer, 1.0 M, pH 8.0, Molecular Biology Grade
648314-100ML
Tris Buffer, 100 mM, pH 7.4, Molecular Biology Grade
648315-100ML
Tris, Hydrochloride, Molecular Biology Grade
648317-100GM
648317-1KG
Tris, Hydrochloride, ULTROL Grade
648313-250GM
648313-500GM
648313-1KG
Tris, ULTROL Grade
648311-1KG
648311-5KG

100 g
1 kg
100 mL
10 1L
10 1L
25 g
250 g
1 kg
10 kg
100 g
100 g
1 kg
100 g
250 g
1 kg
250 g
1 kg
100 g
1 kg
5 kg
500 g
1 kg
250 g
1 kg
500 g
1 kg
250 g
1 kg
2 pack
2 pack
1L
1 kg
2 pack
100 mL
100 g
1 kg
25 g
250 g
10 kg
2 pack
2 pack
100 g
1 kg
1L
500 g
2.5 kg
100 mL
100 mL
100 g
1 kg
250 g
500 g
1 kg
1 kg
5 kg

31

104936

GR

1.04936.0500

100317

GR

1.00317.1000

1L

100244

GR

1.00244.0500

500 g

2 GR

1.06448.0500

500 g

3570-500GM

500 g

106448
3570

3570-1KG
357002

104201

GR

357002-100GM

500 g

1 kg
100 g

357002-1KG

1 kg

1.04201.0100

100 g

1.04201.0250

250 g

1.04201.1000

1 kg

1.04201.5000

5 kg

100062

GR

1.00062.1000

1L

106267

3 GR

1.06267.0500

500 g

106268

GR

1.06268.0250

250 g

106498

GR

1.06498.0500

500 g

106329

GR

1.06329.0500

500 g

106393

) GR

1.06393.1000

1 kg

108382

( GR

1.08382.0100

100 g

1.08382.0500

500 g

1.08382.1000

1 kg

1.08382.2500

2.5 kg
250 g

648309

648309-250GM

648310

648310-500GM

500 g

648310-2.5KG

2.5 kg

648311
106586

( ULTROL

648311-1KG

1 kg

648311-5KG

5 kg

1.06586.0500

500 g
500 g

10658

2 GR

1.06580.0500

106575

7 GR

1.06575.1000

1 kg

106579

12 GR

1.06579.0500

500 g

106346

1 GR

1.06346.0500

500 g

pH
NIST pH
0.003)
0.015)
0.02
http://www.merck.co.jp/request/

32

CalbiochemNovagen

http://bio.merck.co.jp/

Calbiochem

1500
37

http://www.merck.co.jp/request/

Calbiochem

(Oncogene Research Products)

1 2000
ELISA

http://www.merck.co.jp/request/

 18 10 20 BIO03-0610-5000