NuPAGE® SDS-PAGE Gel Protocols

For complete protocols for sample preparation, buffer preparation, electrophoresis, staining, and blotting using the NuPAGE® Electrophoresis System, consult the NuPAGE® Technical Guide.

NuPAGE® Technical Guide

Protocol for 1D SDS Gel Electrophoresis of Proteins with NuPAGE® Novex® Pre-Cast Gels

Get detaied protocol information for one-dimensional SDS gel electrophoresis of proteins with NuPAGE® Novex® Pre-Cast Gels. The protocol includes information on:

  • Materials & Ordering Information
  • Preparing Running Buffer
  • Preparing Sample
  • Using ZOOM® Gels
  • Calibrating Protein Molecular Weight
  • Troubleshooting

    Protocol

Turbo Protocol

Discover the Power and Performance of Novex® NuPAGE® Precast Gel
An optional accelerated process is available for achieving excellent separation and resolution in only 25 minutes when using NuPAGE® Novex® Bis-Tris Gels with MES SDS Running Buffer (Figure 1). The voltage is set at 250 V constant with an expected current of 130–150 mA per gel, and a run time of 25 minutes. With the NuPAGE® Turbo protocol, you get:

  • Advanced technologies that allow an accelerated process
  • Excellent separation and resolution in only 25 minutes
NuPAGE Turbo Protocol


Figure 1. NuPAGE® Turbo protocol. Gels run even faster with the Turbo protocol — only 25 minutes with no band distortion at 250V. Novex® NuPAGE® 4–12% Bis-Tris gels were loaded with the following samples and run with MES SDS running buffer using the standard protocol at 200 V for 40 minutes (right) and the accelerated protocol at 250 V for 25 minutes (left). The gels were stained with Coomassie® R-250. Lanes 1, 3, 5, 7, 9: 5 µL SeeBlue® Plus2 Prestained Standard; Lanes 2, 4, 6, 8, 10: 5µL Mark 12™ Unstained Standard; Lysate: 10 µg E. Coli lysate.

NuPAGE® SDS-PAGE Gel Run Comparison

Gel / Run ConditionRun Time/Expected CurrentComment
NuPAGE® Mini-Gel at 200V
(MES Buffer + Antioxidant) 1 or 2 gels		35 minutes
110-125mA per gel start / 70-80mA per gel end		Optimize separation and resolution
NuPAGE® Mini-Gel at 250V
(MES Buffer + Antioxidant) 1 or 2 gels		24 minutes
130-150mA per gel start / 130-140mA per gel end		No significant band distortion when running 1 or 2 gels at a time
NuPAGE® Midi-Gel at 200V
(MES Buffer + Antioxidant) 1 – 4 gels		40 minutes
250-270mA per gel start / 150-170mA per gel end		Optimize separation and resolution
NuPAGE® Midi-Gel at 250V
(MES Buffer + Antioxidant) 1-4 gels		25 minutes
270-290mA per gel start / 270-290mA per gel end		No significant band distortion when running 4 gels at a time
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
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