Bio-Rad PROTEAN® Plus Hinged Spacer Plates and Combs - Manual (Page 12)

Fig. 7.5. Convex gradient gel.
Section 8
Protocols
8.1 Preparation of Stock Solutions for SDS-PAGE Slab Gels
(Laemmli* buffer system)
A. Acrylamide/Bis (30%T, 2. 67%C)
Acrylamide 146.0 g
N'N'-BIS-methylene-acrylamide 4.0 g
(Preweighed Acrylamide/Bis 37.5:1 mixture may be substituted)
Distilled water to 500 ml. Filter and store at 4 °C in the dark. Maximum shelf life under
these conditions is 30 days.
B. 1.5 M Tris-HCl, pH 8.8
54.45 g Tris base
150 ml distilled water
Adjust to pH 8.8 with 1 N HCl. Distilled water to 300 ml. Store at 4 °C.
C. 0.5 M Tris-HCl, pH 6.8
6 g Tris base
60 ml distilled water
Adjust to pH 6.8 with HCl. Distilled water to 100 ml. Store at 4 °C.
D. 10% (w/v) SDS
Dissolve 10 g SDS in water with gentle stirring. Distilled water to 100 ml.
E. 10% Ammonium Persulfate (w/v)
100 mg ammonium persulfate. Distilled water to 1 ml. Make fresh daily.
10
% Acrylamide 20
15
10
Amount delivered (%)
25 50 75 100