5. Operation
B. Sample buffer
0.050 mol/l Tris-HAc pH 7.5
Dissolve 0.3 g Tris in 40 ml distilled water. Carefully adjust to pH 7.5 with
HAc (approximately 0.14 ml). Make up to 50 ml with distilled water. Add
0.4 g SDS and a few grains of Bromophenol Blue. Immediately before use
add 40 mg of DTT.
C. Fixing solution
Ethanol
400 ml
Acetic acid, HAc
100 ml
Make up to 1000 ml with distilled water.
D. Incubation solution
Ethanol
75 ml
Sodium acetate
17.00 g
Glutaraldehyde (25% w/v)
1.25 ml
Sodium thiosulphate, Na
2
S
2
O
3
x 5H
2
O
0.50 g
Make up to 250 ml with distilled water.
E. Silver solution
Silver nitrate
0.25 g
Formaldehyde
50 µl
Make up to 250 ml with distilled water.
F. Developing solution
Sodium carbonate
6.25 g
Formaldehyde
25 µl
Make up to 250 ml with distilled water.
G. Stop solution
EDTA-Na
2
x 2H
2
O
3.65 g
Make up to 250 ml with distilled water.
H. Preserving solution
Glycerol (87% w/w)
25 ml
Make up to 250 ml with distilled water.
I. Destaining solution
Ethanol
250 ml
Acetic acid
80 ml
Make up to 1000 ml with distilled water.
K. Coomassie solution
PhastGel Blue R
1 tablet
Make up to 400 ml with destaining solution.
Heat to 60 °C, stirring constantly, and filter before use.
L. Preserving solution
Glycerol (87% w/w)
25 ml
Make up to 250 ml with destaining solution.
N. Fixing solution
Trichloroacetic acid
100 g
Make up to 500 ml with distilled water.
47
5.9 Stock
solutions
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