Protein Analysis - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 168

Fig. 1. The setup of the polyacrylamide gel caster.

3. Typically, 10 ml of the resolving gel mixture is needed to cast
at least one gel. The recipes for the different percentage cell
mixture is as follows:

30%
Acrylamide
mix (ml)

1.5 M Tris
(pH 8.8) (ml)

10% SDS
( m l)

H 2 O
(ml)

10% APS
( m l)

TEMED
( m l)

8% 2.7 2.5 100 4.6 100 10

10% 3.3 2.5 100 4.0 100 10

12% 4.0 2.5 100 3.3 100 10

15% 5.0 2.5 100 2.3 100 10

4. Pour the resolving gel mixture into the space between the
two glass plates less than 1.5 cm below the top of the longer
glass plate. Then layer 100% ethanol onto the gel mixture
interface to minimize its contact to air and facilitate the
polymerization.
5. After the resolving gel is set, the ethanol is pour out and the
interface is rinse once with water. Drain the excess water before
pouring the stacking gel mixture on top of the resolving gel.
The recipe of the stacking gel mixture (4 ml) is as follows: