Hoefer SE900 - Manual (Page 32)

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6. 0.375 M Tris-Cl, 0.1% SDS, pH 8.8
(Resolving gel overlay, 100 ml)
1.5 M Tris-Cl, pH 8.8
(Solution #2) 0.375 M 25 ml
10% SDS (Solution #4) 3.5 mM 1 ml
Deionized H
2
O to 100 ml.
Store at room temperature.
— OR —
Water-saturated n-butanol
Shake n-butanol and deionized H
2
O in a separatory funnel. Remove the
aqueous (lower) phase. Repeat this procedure several times. Use the
upper phase.
Store at room temperature.
7. 10X Electrophoresis Buffer
0.25 M Tris, 1.92 M Glycine, 1.0% SDS
(10X Electrophoresis buffer, 5.0 liters)
Add powders slowly to ~4 liters deionized water while stirring.
Tris
(FW 121.1) 0.25 M 151.2 g
Glycine (FW 75.07) 1.92 M 720.6 g
SDS (FW 288.4) 35 mM 50.0 g
Deionized H
2
O to 5.0 liters.
The pH of this buffer is approximately 8.3. Do not adjust pH.
Store room temperature for up to 2 months.
8. 1X Electrophoresis Buffer
0.025 M Tris, 0.192 M Glycine, 0.1% SDS
(1X Electrophoresis buffer, 12.0 liters)
10X Electrophoresis Buffer (Solution #7)
1200 ml
Deionized H
2
O 10.8 l
This can be prepared in the separation tank. Add 10X stock, water then the
PAGE rack. Gently raise and lower the PAGE rack to aid in mixing then allow
the recirculation system to mix the buffer well before use.