10
Table 1.
ENDURO VE10
Total Gel volume for a 1mm thick gel.
For different thicknesses of gel, multiple the below amounts by the spacer
thickness.
Single – one gel, one dummy plate
7.5 mL
Double – two gels
15 mL
Using a Triple Plate sandwich – 4 gels
30 mL
Gel Selection:
Care should be taken when selecting the pore size of the gel to be used. These
formulas are for Tris- glycine-SDS gels
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 %
60 - 220 KD
7.5 %
45 - 120 KD
10 %
25 - 75 KD
12%
14.4 – 65 KD
15 %
6.5 -45 KD
17.5%
5.5 – 30 KD
3. Using the stock solution provided in the appendix prepare gel solutions as per
tables below. First mix the ddi Water, 30% Acrylamide solution and the 4x TRIS-
SDS solutions. After mixing, degas for 5 minutes to remove free oxygen (which
will inhibit polymerization).
4. To the above solution add the ammonium persulfate and TEMED and mix gently
to avoid air bubbles.
