Electrophoresis buffer, Coomassie stain solution, Destain solution i – Hoefer SE400 User Manual
Page 40: Destain solution ii, Cross-linking solution, Dtt (dithiothreitol) solution
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8. Electrophoresis buffer
(0.025 M Tris, 0.192 M glycine, 0.1% SDS, pH 8.3,
5.0 liters)
Tris (FW 121.1)
0.025 M
15.1 g
Glycine (FW 75.07)
0.192 M
72.1 g
SDS (FW 288.4)
3.5 mm
5.0 g
Deionized H
2
O
to 5.0 liters
The pH of this buffer is approximately 8.3. Do not adjust pH.
Up to 20 liters can be prepared and stored for up to 2
months.
9. Coomassie stain solution
(0.025% Coomassie Blue R-250, 40% Methanol,
7% Acetic acid, 2 liters)
Coomassie Blue R-250 (FW 826)
0.3 mm
0.5 g
Methanol (Stir until dissolved)
40% v/v
800.0 ml
Acetic acid
7% v/v
140.0 ml
Deionized H
2
O
to 2.0 liters
10. Destain solution I
(40% methanol, 7% acetic acid, 1 liter)
Methanol
40% v/v
400.0 ml
Acetic acid
7% v/v
70.0 ml
Deionized H
2
O
to 1.0 liter
11. Destain solution II
(7% acetic acid, 5% methanol)
Methanol
5% v/v
50.0 ml
Acetic acid
7% v/v
70.0 ml
Deionized H
2
O
to 1.0 liter
12. Cross-linking solution
(10% glutaraldehyde)
20 ml of 50% glutaraldehyde stock
Distilled water to 100 ml.
13. DTT (dithiothreitol) solution
(5 µg/ml)
5 mg DTT
Bring to 1 L with ddH
2
O.
Caution! Glutaraldehyde should
only be handled in a fume hood.