Gel Selection - VWR shiroGEL Vertical 20 Manual De Instrucciones

Gel Selection:

Care should be taken when selecting the pore size of the gel to be used.
The pore size or % of gel determines the resolving ability given different sizes of
protein.
See Table 2 below which details which percentage of gel to use to separate the
sizes of proteins indicated.
Table 2.
Acrylamide Percentage Separating Resolution
5 %
7.5 %
10 %
12%
15 %
17.5%
3. Prepare gel solutions as per tables below. These give the volumes of solutions
from the standard stock solutions. These should be gently mixed avoiding
generation of bubbles which will inhibit polymerization by removing free radicals.
Table 3: Preparation of the separating gel solution for two 20 x 20cm shiroGEL
VERTICAL 20 gels using 1 mm spacers.
Solution
Distilled Water
30 % Stock Acrylamide
Solution
4 X Resolving Tris
Solution
10 % Ammonium
Persulphate
60 - 220 KD
30 - 120 KD
20 - 75 KD
17 – 65 KD
15 -45 KD
12 – 30 KD
5 % 7.5%
41ml 35.25ml
11.7ml 17.6ml
17.6ml 17.6ml
700µl 700µl
17
10 % 12%
29.6ml 24.7ml 17.6ml
23.5ml 28.2ml 35.25ml
17.6ml 17.6ml 17.6ml
700µl 700µl
15 % 17.5%
11.7ml
41.1ml
17.6ml
700µl 700µl