Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 23)

7.4 Zymogram Buffers
Running buffer 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 Working Concentration 2X Stock
62.5 mM Tris-HCl, pH 6.8 0.5 M Tris-HCl, pH 6.8 1.25 ml
4% SDS 10% SDS 4.0 ml
25% glycerol Glycerol 2.5 ml
0.01% Bromophenol Blue 1% Bromophenol Blue 0.1 ml
Deionized water 2.15 ml
10 ml
7.5 Sample Preparation
Determine the appropriate protein concentration of your sample based on the detection method and load
volume used. (See section 10.4 for approximate stain sensitivities.) Dilute 1 part sample buffer with 1 part
Zymogram sample buffer. Dry samples can be dissolved directly in sample buffer. Do not heat.
7.6 Running Conditions
Power conditions 100 V constant
Starting current:
10–15 mA/gel
Final current: 6 mA/gel
Run time 90 min
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