Biomedical Engineering Reference
In-Depth Information
Table 2.1.
Recipes for casting gels with homogeneous acrylamide con-
centration (L
AEMMLI
system)
%T = 5.0
a
7.5
a
10.0
a
12.0
b
15.0
b
20.0
b
Stacking gel
c
Solution
(ml/10 ml)
A
1.22
1.83
2.43
-
-
-
1.10
A
-
-
-
2.97
3.55
4.75
-
B
3.75
3.75
3.75
3.75
3.75
3.75
-
C
-
-
-
-
-
-
2.50
D
0.05
0.05
0.05
0.05
0.05
0.05
0.05
E
0.10
0.10
0.10
0.10
0.10
0.10
0.10
Add H
2
O to 9.90 ml
F
0.10
0.10
0.10
0.10
0.10
0.10
0.10
a
%C = 2.68
b
%C=5.0
c
%T = 4.5, %C = 2.68
Table 2.2.
Recipes for casting gels with acrylamide concentration gradi-
ent 5 → 15 and 5 →
20
%, respectively (L
AEMMLI
system)
Light solution
Dense solution
%T = 5.0%
%T = 15.0
%T = 20.0
%C=2.68
%C=5
Solution
(ml/10 ml; total volume 20 ml)
A
1.22
-
-
A
-
3.55
4.75
B
3.75
3.75
3.75
D
0.05
0.05
0.05
E
0.05
0.05
0.05
H
2
O to 9.95
Add glycerol to 9.95 ml
F
0.05
0.05
0.05
a
For gradients with other %T and %C, amounts of Soln. A and
A
, respectively, are given in Table 2.1
gel. The electrode buffer may be used for several times if pH is not
shifted and if it is not contaminated by constituents of the samples.
Important!
Remove the comb
after
filling the electrophoresis cham-
bers with electrode buffer and remove air bubbles within the slots.
If the sample buffer G is used for dissolving precipitated or
Sample preparation
lyophilized samples (sometimes such samples are difficult to dis-
solve in Soln. G
), some glucose, sucrose, or glycerol and up to 5%
of sample volume of saturated bromophenol blue in Soln. G may
be added to ease sample application.
If potassium ions are present within the sample, potassium
dodecylsulfate precipitates and dodecylsulfate ion concentration is
