Making up the Solutions:
The following tables display the components of the stock solutions used in our experiments, including the relative amounts of each component needed.
Wash Buffer
|
|
1 gel |
2 gel |
|
50 mM Tris-HCl (pH 7.5) |
0.61 g in 50 ml ddH2O |
1.22 g in 100 ml ddH2O |
|
10 % Triton X-100 |
25 ml |
50 ml |
|
ddH2O |
25 ml |
50 ml |
Incubation Buffer
|
|
1 gel |
2 gel |
|
Wash Buffer |
25 ml |
50 ml |
|
50 mM MgCl2 |
0.12 g |
0.24 g |
|
62.5 mM CaCl2 |
0.18 g |
0.36 g |
0.25 % Coomassie Blue Stain
|
|
100 ml |
200 ml |
500 ml |
1 L |
|
Coomassie Blue R-250 |
0.25 g |
0.50 g |
1.25 g |
2.5 g |
|
Methanol
|
40 ml |
80 ml |
200 ml |
400 ml |
|
ddH2O
|
50 ml |
100 ml |
250 ml |
500 ml |
|
Acetic Acid
|
10 ml |
20 ml |
50 ml |
100 ml |
Coomassie Blue Destain
|
|
100 ml |
200 ml |
500 ml |
1 L |
|
Methanol |
50 ml |
100 ml |
250 ml |
500 ml |
|
ddH2O
|
40 ml |
80 ml |
200 ml |
400 ml |
|
Acetic Acid |
10 ml |
4 ml |
10 ml |
10 ml |
Zymography Samples
|
Sample |
Protein Concentration |
Protein Volume |
Water (ml) |
SB (ml) |
Total Protein (mg) |
|
1-1 |
0.87 |
5.1 |
4.9 |
10 |
3.3 |
|
1-7 |
1.06 |
4.2 |
5.8 |
10 |
3.3 |
|
2-1 |
1.33 |
3.3 |
6.7 |
10 |
3.3 |
|
2-7 |
1.30 |
3.4 |
6.6 |
10 |
3.3 |
|
3-1 |
2.17 |
2.0 |
8.0 |
10 |
3.3 |
|
3-7 |
2.86 |
1.5 |
8.5 |
10 |
3.3 |
|
4-1 |
0.44 |
10 |
0 |
10 |
3.3 |
|
4-7 |
0.71 |
6.2 |
3.8 |
10 |
3.3 |
|
5-1 |
0.93 |
4.7 |
5.3 |
10 |
3.3 |
|
5-7 |
1.29 |
3.4 |
6.6 |
10 |
3.3 |
|
6-1 |
2.07 |
2.1 |
7.9 |
10 |
3.3 |
|
6-7 |
1.67 |
2.6 |
7.4 |
10 |
3.3 |
|
7-1 |
0.53 |
8.3 |
1.7 |
10 |
3.3 |
|
7-7 |
0.63 |
7.0 |
3.0 |
10 |
3.3 |
|
8-1 |
1.17 |
3.8 |
6.2 |
10 |
3.3 |
|
8-7 |
1.13 |
3.9 |
6.1 |
10 |
3.3 |
|
9-1 |
1.28 |
3.4 |
6.6 |
10 |
3.3 |
|
9-7 |
1.38 |
3.2 |
6.8 |
10 |
3.3 |
1- wet white rice 4- KCl white rice 7- NaCl white rice
2- wet brown rice 5- KCl brown rice 8- NaCl brown rice
3- wet brown basmati rice 6- KCl brown basmati rice 9- NaCl brown basmati rice