Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 14)

3.4 SDS-PAGE Buffers
Running Buffer 1X Working Concentration 10x Stock
25 mM Tris Tris base 15.0 g
192 mM glycine Glycine 72.0 g
0.1% SDS SDS 5.0 g
to 500 ml with deionized water
Note: running buffer should be
~ pH 8.3. Do not adjust the pH.
Sample Buffer 2X Working Concentration 2X Stock
62.5 mM Tris-HCl, pH 6.8 0.5 M Tris-HCl, pH 6.8 1.0 ml
2% SDS 10% (w/v) SDS 1.6 ml
25% glycerol Glycerol 2.0 ml
0.01% Bromophenol Blue 1.0% Bromophenol Blue 0.08 ml
5% 2-mercaptoethanol 2-Mercaptoethanol 0.4 ml
or 350 mM DTT (added fresh) Deionized water 2.92 ml
8.0 ml
3.5 Sample Preparation
Determine the appropriate protein concentration of your sample based on the detection method and load
volume used. (See section 10.1 for approximate stain sensitivities.) Dilute 1 part sample with 1 part sample
buffer (see section 3.4) and heat at 95ºC for 5 min.
3.6 Running Conditions
Power conditions 200 V constant
Starting current:
50 mA/gel
Final current: 30 mA/gel
Run time 35 min
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