Bio-Rad Criterion™ XT Tris-Acetate Precast Gels - Manual (Page 13)

3.5 Sample Preparation
Determine the appropriate protein concentration of your sample based on the detection method and load
volume used. (See section 4.1 for approximate stain sensitivities.) XT sample buffer is a 4x concentrate and
can be used with both dilute and concentrated samples. Refer to the sample preparation guide below:
Sample Preparation Guide
SDS-PAGE Native-PAGE
25 µl XT sample buffer 50 µl Native sample buffer
5 µl XT reducing agent x µl sample
x µl sample
Make up to 100 µl with ddH20 Make up to 100 µl with ddH2O
Heat sample at 95°C for 5 min. Do not heat sample
3.6 Running Conditions
Bis-Tris Bis-Tris Tris-Acetate Tris-Acetate
Gel type (for SDS-PAGE) (for SDS-PAGE) (for SDS-PAGE) (for Native-PAGE)
Running buffer XT MOPS XT MES XT Tricine Tris/Glycine
Power conditions 200 V constant 200 V constant 150 V constant 200 V constant
Run time 60 min 45 min 65 min 75 min
Starting current 165–175 mA/gel 185–200 mA/gel 170–180 mA/gel 70–80 mA/gel
Final current 60–70 mA/gel 90–110 mA/gel 85–95 mA/gel 25–35 mA/gel
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