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® precast gels

Get detailed protocol information for one-dimensional SDS gel electrophoresis of proteins with NuPAGE® Novex® precast 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
photo of stained gels showing results when using Turbo protocol


Figure 1. NuPAGE® Turbo protocol. Gels run even faster with the Turbo protocol—only 25 minutes with no band distortion at 250 V. 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 condition Run time/expected current Comment
NuPAGE® Mini gel at 200 V
(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 250 V
(MES Buffer + Antioxidant) 1 or 2 gels
24 minutes
130–150 mA per gel start / 130–140 mA per gel end
No significant band distortion when running 1 or 2 gels at a time
NuPAGE® Midi gel at 200 V
(MES Buffer + Antioxidant) 1–4 gels
40 minutes
250–270 mA per gel start / 150–170 mA per gel end
Optimize separation and resolution
NuPAGE® Midi gel at 250 V
(MES Buffer + Antioxidant) 1–4 gels
25 minutes
270–290 mA per gel start / 270–290 mA per gel end
No significant band distortion when running 4 gels at a time