User Manual

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Table A.1. Running conditions for SDS-PAGE in the Criterion cell. Do not run different gel formulations at the same time.
TGX Tris-HCl Bis-Tris Tris-Acetate
Running buffer Tris/glycine/SDS Tris/glycine/SDS XT MOPS XT MES XT Tricine
Standard Conditions
Power conditions 200 V constant 200 V constant 200 V constant 200 V constant 150 V constant
Expected current (per gel)
Initial 55–80 mA 90–120 mA 165–175 mA 185–200 mA 170–180 mA
Final 33–43 mA 35–55 mA 60–70 mA 90–110 mA 85–95 mA
Run time 42–45 min 50–55 min 60 min 45 min 65 min
High Voltage (Rapid) Conditions
Power conditions 300 V constant — — — —
Expected current (per gel)
Initial 89–135 mA — — — —
Final 66–99 mA — — — —
Run time 20–26 min — — — —
Table A.2. Standard transfer (blotting) conditions.
Method Membrane Time Voltage
Tank blotting Nitrocellulose (0.45 μm) 20–60 min 100 V
1
PVDF (0.2 μm)
Semi-dry blotting Nitrocellulose (0.45 μm) 20–60 min 25 V
PVDF (0.2 μm)
1
For previously optimized protocols for proteins <30 kD, reduce transfer time by 20%.