Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 21)

6.4 IEF Buffers
Running buffer 1x Cathode Buffer 5x Cathode Buffer
20 mM lysine (free base) Lysine (free base) 14.50 g
20 mM arginine (free base) Arginine (free base) 17.42 g
to 1 L with deionized water
1x Anode Buffer 10x Anode Buffer
7mM% phosphoric acid Phosphoric acid 4.2 ml
to 1 L with deionized water
Sample Buffer 50% glycerol
6.5 Sample Preparation
Determine the appropriate protein concentration of your sample based on the detection method and load
volume used. (See section 10.3 for approximate stain sensitivities.) Dilute 1 part sample with 1 part sample
buffer.
6.6 Running Conditions
Power conditions Stepwise
100 V constant
60 min
250 V constant 60 min
500 V constant 30 min
Starting current: 5–15 mA/gel
Final current: 5–15 mA/gel
Run time 150 min
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