Bio-Rad Mini-PROTEAN® Tetra Cell - Manual (Page 26)

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4.3 Discontinuous Native PAGE (Ornstein-Davis) Stock
Solutions and Buffers
1. Acrylamide/Bis (30%T, 2.67%C)
87.6 g acrylamide (29.2 g/100 ml)
2.4 g N’N’-bis-methylene-acrylamide (0.8 g/100 ml)
Make to 300 ml with deionized water. Filter and store at 4°C in
the dark (30 days maximum).
Or, use:
Preweighed acrylamide/bis, 37.5:1 mixture
(Bio-Rad catalog #161-0125, 150 g)
30% acrylamide/bis solution, 37.5:1 mixture
(Bio-Rad catalog #161-0158, 500 ml)
(Bio-Rad catalog #161-0159, 2 x 500 ml)
2. 1.5 M Tris-HCl, pH 8.8
27.23 g Tris base (18.15 g/100 ml)
80 ml deionized water
Adjust to pH 8.8 with 6 N HCl. Bring total volume up to 150
ml with deionized water and store at 4°C. Alternatively, 1.5 M
Tris-HCl, pH 8.8 (1 L) premixed buffer can be used (Bio-Rad
catalog #161-0798).
3. 0.5 M Tris-HCl, pH 6.8
6 g Tris base
60 ml deionized water
Adjust to pH 6.8 with 6 N HCl. Bring total volume up to 100
ml with deionized water and store at 4°C. Alternatively, 0.5 M
Tris-HCl, pH 6.8 (1 L) premixed buffer can be used (Bio-Rad
catalog #161-0799).
4. Sample Buffer
5.55 ml deionized water
1.25 ml 0.5 M Tris-HCl, pH 6.8
3.0 ml glycerol
0.2 ml 0.5% (w/v) Bromophenol Blue
10.0 ml Total volume
Store at room temperature.
Use: Dilute the sample at least 1:2 with sample buffer.