Bio-Rad Mini-PROTEAN® Tetra Cell - Manual (Page 23)

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Section 4
Reagent Preparation and Stock Solutions
4.1 Volumes Required Per Gel
The volumes listed are required to completely fill a gel cassette.
Amounts may be adjusted depending on the application (with or
without comb, with or without stacking gel, etc.).
Gel Thickness (mm) Volume (ml)
0.5 2.8
0.75 4.2
1.0 5.6
1.5 8.4
Note: 10 ml of monomer solution is sufficient for two stacking gels
of any thickness.
4.2 SDS-PAGE (Laemmli) Buffer System 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 (30%T, 2.67% C)
(Bio-Rad catalog #161-0125, 150 g)
30% acrylamide/bis solution, 37.5:1 mixture (30%T, 2.67% C)
(Bio-Rad catalog #161-0158, 500 ml)
(Bio-Rad catalog #161-0159, 2 x 500 ml)
2. 10% (w/v) SDS
Dissolve 10 g SDS in 90 ml water with gentle stirring and
bring to 100 ml with deionized water. Alternatively, 10% SDS
solution (250 ml) can be used (Bio-Rad catalog #161-0416).
3. 1.5 M Tris-HCl, pH 8.8
27.23 g Tris base (18.15 g/100 ml)
80 ml deionized water