Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 19)

5.4 Tris-Tricine/Peptide Buffers
Running buffer Working Concentration 10x Stock
100 mM Tris Tris base 60.55 g
100 mM Tricine Tricine 89.60 g
0.1% SDS SDS 5.0 g
to 500 ml with deionized water
Note: Tricine running buffer should be
~ pH 8.25. Do not adjust the pH.
Sample Buffer Working Concentration 2X Stock
200 mM Tris-HCl, pH 6.8 1.0 M Tris-HCl, pH 6.8 2.0 ml
2% SDS 10% SDS 2.0 ml
40% glycerol Glycerol 4.0 ml
0.04% Coomassie Blue G-250 0.5% Coomassie Blue G-250 0.8 ml
2% 2-mercaptoethanol 2-Mercaptoethanol 0.2 ml
or 350 mM DTT (Added fresh) Deionized water 1.0 ml
10.0 ml
5.5 Sample Preparation
Determine the appropriate protein concentration of your sample based on the detection method and load
volume used. (See section 10.2 for approximate stain sensitivities.) Dilute 1 part sample with 1 part sample
buffer and heat at 95ºC for 5 min.
5.6 Running Conditions
Power Conditions 100 V constant
Starting current:
30–35 mA/gel
Final current: 15–20 mA/gel
Run Time 100 min
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