Acids and alkalis.
| Name:
|
Formula:
|
Mw
|
% (w/w)
|
[M]
|
g in 1L
of subst.
|
p [g/ml]
|
ml/L for
1M sol.
|
| Sodium hydroxide
|
NaOH
|
40
|
50%
|
19.1
|
763
|
1.53
|
52.4
|
| 30.1%
|
10.0
|
400
|
1.329
|
100
|
| 10%
|
2.75
|
111
|
1.11
|
363.6
|
|
Potassium hydroxide
|
KOH
|
56.1
|
50%
|
13.5
|
757
|
1.52
|
74.1
|
| 23.06%
|
5.0
|
280.6
|
1.217
|
200
|
| 10%
|
1.94
|
109
|
1.09
|
515.5
|
| Ammonium hydroxide
|
NH4
OH
|
35.0
|
28%
|
14.8
|
251
|
0.898
|
67.6
|
| Acetic acid, glacial
|
CH3
COOH
|
60.05
|
99.5%
|
17.4
|
1045
|
1.05
|
57.5
|
| Acetic acid
|
36%
|
6.27
|
376
|
1.045
|
159.5
|
| Formic acid
|
HCOOH
|
46.02
|
90%
|
23.4
|
1080
|
1.20
|
42.7
|
| Hydrochloric acid
|
HCl
|
36.5
|
36%
|
11.6
|
424
|
1.18
|
86.2
|
| 10%
|
2.9
|
105
|
1.05
|
344.8
|
| Nitric acid
|
HNO3
|
63.02
|
71%
|
15.99
|
1008
|
1.42
|
62.5
|
| 67%
|
14.9
|
938
|
1.40
|
67.1
|
| 61%
|
13.3
|
837
|
1.37
|
75.2
|
| Perchloric acid
|
HClO4
|
100.5
|
70%
|
11.65
|
1172
|
1.67
|
85.8
|
| 60%
|
9.2
|
923
|
1.54
|
108.7
|
| Phosphoric acid
|
H3
PO4
|
98.0
|
85%
|
18.1
|
1445
|
1.71
|
55.2
|
| Sulfuric acid
|
H2
SO4
|
98.1
|
96%
|
18.0
|
1766
|
1.84
|
55.6
|
|
NaOH
Mw=40g/M; (store at NT).
|
|
Conc.
|
Stock
|
%(w/w)
|
50ml
|
200ml
|
300ml
|
|
| NaOH
|
10M
|
40g/M
|
30.1
|
20g
|
80g
|
120g
|
|
| H2
O
|
|
mQ
|
69.90
|
46.45ml
|
185.8ml
|
278.7ml
|
|
|
10M: 30.10%; p=1.329.
|
|
Conc.
|
Stock
|
%(w/w)
|
50ml
|
150ml
|
200ml
|
|
| NaOH
|
1M
|
10M
|
12.87
|
5ml
6.645g
|
15ml
19.94g
|
20ml
26.58g
|
|
| H2
O
|
|
mQ
|
87.13
|
45ml
|
135ml
|
180ml
|
|
|
- it is better to use plastic bottles for storage, because the alkali slightly solubilize the glass.
KOH
Mw=56.11g/M; (store at NT).
|
|
Conc.
|
Stock
|
%(w/w)
|
10ml
|
50ml
|
100ml
|
150ml
|
|
| KOH
|
5M
|
56.11g/M
|
23.05
|
2.806g
|
14.03g
|
28.06g
|
42.08g
|
|
| H2
O
|
|
mQ
|
76.95
|
9.363g
|
46.81g
|
93.63g
|
140.4g
|
|
|
5M: 23.06%; p=1.217.
- it is better to use plastic bottles for storage, because the alkali slightly solubilize the glass.
TCA
CCl3
CO2
H Mw=163.39g/M,(store at NT, in the dark, under the fume hood).
|
|
Conc.
|
%(w/w)
|
5ml
|
10ml
|
25ml
|
50ml
|
|
| TCA
|
100% (w/v)
|
68.78
|
5.0g
|
10.0g
|
25.0g
|
50.0g
|
|
| H2
O
|
mQ
|
31.22
|
2.27ml
|
4.54ml
|
11.35ml
|
22.7ml
|
|
|
Detergents.
| Detergent
|
Tmelt
|
Mw [Da]
|
CMC
|
| monomer
|
micelle
|
%(w/v)
|
M
|
| Anionic
|
| SDS
|
206
|
288
|
18,000
|
0.23
|
8.0 x 10-3
|
| Cholate
|
201
|
430
|
4,300
|
0.60
|
1.4 x 10-2
|
| Deoxycholate
|
175
|
432
|
4,200
|
0.21
|
5.0 x 10-3
|
| Cationic
|
| C16
TAB
|
230
|
365
|
62,00
|
0.04
|
1.0 x 10-3
|
| Amphoteric
|
| LysoPC
|
-
|
495
|
92,000
|
0.0004
|
7.0 x 10-6
|
| CHAPS
|
157
|
615
|
6,150
|
0.49
|
1.4 x 10-3
|
| Zwittergent 3-14
|
-
|
365
|
30,000
|
0.011
|
3.0 x 10-4
|
| Nonionic
|
| Octyl glucoside
|
105
|
292
|
8,000
|
0.73
|
2.3 x 10-2
|
| Digitonin
|
235
|
1,229
|
70,000
|
-
|
-
|
| C12
E8
|
-
|
542
|
65,000
|
0.005
|
8.7 x 10-5
|
| Lubrol PX
|
-
|
582
|
64,000
|
0.006
|
1.0 x 10-4
|
| Triton X-100
|
-
|
650
|
90,000
|
0.021
|
3.0 x 10-4
|
| Nonident P-40
|
-
|
603
|
90,000
|
0.017
|
3.0 x 10-4
|
| Tween 80
|
-
|
1,310
|
76,000
|
0.002
|
1.2 x 10-5
|
|
CMC - critical micelle concentration.
| Detergent
|
1
|
2
|
3
|
4
|
5
|
6
|
7
|
8
|
9
|
10
|
11
|
12
|
1
- Strongly denaturing;
2
- Dializable;
3
- Ion exchangeable, unsuitable for ion-exchange chromatography;
4
- Complexes ions;
5
- Strong A280
;
6
- Assay interference;
7
- Cold precipitates;
8
- High cost;
9
- Ease of purification;
10
- Radiolabeled;
11
- Definite composition;
12
- Auto-oxidation
|
| Anionic
|
| SDS
|
+
|
+
|
+
|
+
|
-
|
-
|
+
|
-
|
+
|
+
|
+
|
-
|
| Cholate
|
-
|
+
|
+
|
+
|
-
|
-
|
-
|
-
|
+
|
+
|
+
|
-
|
| Deoxycholate
|
-
|
+
|
+
|
+
|
-
|
-
|
+
|
-
|
+
|
+
|
+
|
-
|
| Cationic
|
| C16
TAB
|
+
|
+
|
+
|
-
|
-
|
-
|
+
|
-
|
+
|
-
|
+
|
-
|
| Amphoteric
|
| LysoPC
|
+/-
|
-
|
-
|
-
|
-
|
-
|
-
|
+
|
+/-
|
+
|
+
|
-
|
| CHAPS
|
-
|
+
|
-
|
-
|
-
|
-
|
-
|
+
|
+
|
-
|
+
|
-
|
| Zwittergent 3-14
|
+/-
|
+/-
|
-
|
-
|
-
|
-
|
-
|
+
|
+
|
-
|
+
|
-
|
| Nonionic
|
| Octyl glucoside
|
-
|
+
|
-
|
-
|
-
|
-
|
-
|
+
|
-
|
+
|
+
|
-
|
| Digitonin
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
+
|
+
|
-
|
|
|
| C12
E8
|
-
|
-
|
-
|
+/-
|
-
|
-
|
-
|
+
|
-
|
+
|
-
|
+
|
| Lubrol PX
|
-
|
-
|
-
|
+/-
|
-
|
+/-
|
-
|
-
|
-
|
+
|
-
|
+
|
| Triton X-100
|
-
|
-
|
-
|
+/-
|
+
|
+/-
|
-
|
-
|
-
|
+
|
-
|
+
|
| Nonident P-40
|
-
|
-
|
-
|
+/-
|
+
|
+/-
|
-
|
-
|
-
|
+
|
-
|
+
|
| Tween 80
|
-
|
-
|
-
|
+/-
|
-
|
+/-
|
-
|
-
|
-
|
+
|
-
|
+
|
|
C16
TAB - hexadecyl trimethylammonium bromide;
LysoPC - lysophosphatidylcholine.
* Sodium cholate and sodium deoxycholate are unsoluble at <pH 7.5 or at ionic strength greater then 0.1%. SDS may precipitate below 20o
C;
** Ionic detergents may induce problems with electrophoresis and isoelectric focusing;
*** It is possible to remove by dialysis;
**** Phenol -containing detergents (for example Triton X-100 and NP-40) precipitate during Folin protein assay (but do not interfere with Bradford protein assay).
N-Lauroylsarcosine Na
(store at NT).
|
|
Conc.
|
Stock
|
%(w/w)
|
1ml
|
3ml
|
5ml
|
15ml
|
50ml
|
|
| Sarc
|
10%
|
30%
|
33.93
|
0.333ml
0.342g
|
1.00ml
1.027g
|
1.67ml
1.712g
|
5.00ml
5.14g
|
16.67ml
17.12g
|
|
| H2
O
|
|
mQ
|
66.07
|
0.667ml
|
2.00ml
|
3.33ml
|
10.00ml
|
33.33ml
|
|
|
p=1.017.
SDS
Sodium dodecyl sulfate, sodium lauryl sulfate; [CH3
(CH2
)10
CH2
SO4
]Na; Mw=288.4g/M; (store at NT).
|
|
Conc.
|
Stock
|
%(w/w)
|
100 ml
|
250 ml
|
300 ml
|
400 ml
|
|
| SDS
|
10%(w/v)
|
solid
|
9.82
|
10g
|
25g
|
30g
|
40g
|
|
| H2
O
|
|
mQ
|
90.18
|
91.8 ml
|
229.5 ml
|
275.4 ml
|
367.2 ml
|
|
|
p = 1.018
- weight under the fume hood (and better to wear the mask);
- it is necessary to heat to 60-80o
C. to facilitate solubilization. Check pH. If it differs from neutral (~7.2-7.5) dramatically - adjust by diluted alkali / acid.
Organic solvents.
Phenol
C6
H5
OH; Mw=94.1g/M
p=1.054, tm
= 43, tb
= 182, pKa=10.0
solubility: 6.816
H2
O, unlimited 66
H2
O; unlimited EtOH
Preparation of "acidic" and "neutral" phenol.
a) from the distillation:
- distill phenol under H2
O;
- adjust water to the volume of about 1/10 of phenol phase;
- add 8-hydroxyquinoline to 0.1% (relative to phenol phase) and bMeEtOH (2-mercaptoethanol) to 0.2% (relative to H2
O = 0.02% relative to phenol phase);
---- on this step you obtained the "acidic phenol". Store at -20o
C (~1 year); ----
- add about the same volume of 0.2 M Tris-base to the phenol, mix ~0.5-1h;
- throw away aqueous phase;
- + 0.1V 0.1M Tris-Cl, pH 8.0
- 0.2% bMeEtOH
- mix ~0.5-1h;
- store at 4o
C (in the dark) ~several months.
b) from the good commercial substance:
- saturate phenol by the water (add about 1/5 V );
- then, according to (a), from the p.2.
______________
* For RNA extraction it is better to saturate (just add) "acidic phenol" by the following buffer: (store at 4o
C):
|
|
Conc.
|
Stock
|
100ml
|
|
| AcONa, pH 5.1
|
50mM
|
3M
|
1.67ml
|
|
| EDTA
|
10mM
|
0.5M
|
2.0ml
|
|
| H2
O
|
|
mQ
|
96.0ml
|
|
|
|