The Novex® Tricine Gel System is a modification of the traditional Tris-Glycine gel system that uses a discontinuous buffer system specifically designed for the resolution of low–molecular weight proteins. The Novex® Tricine gel system has significant advantages over traditional Tris-Glycine gel systems for resolving proteins in the molecular weight range of 2–20 kDa.

Advantages of Novex® Tricine gels over the Tris-Glycine gels:

  • Increased resolution of proteins with molecular weights as low as 2 kDa
  • Improved compatibility with  direct protein sequencing applications after transferring to PVDF
  • Minimized protein modification due to the lower pH of the tricine buffering system

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Tris-Glycine gels versus Novex® Tricine gels

In the traditional Tris-Glycine protein gel system, the proteins are stacked in the stacking gel between the highly mobile leading chloride ions (in the gel buffer) and the slower, trailing glycine ions (in the running buffer). These stacked protein bands undergo sieving once they reach the separating gel. However, the resolution of smaller proteins (<10 kDa) is hindered by the continuous accumulation of free dodecyl sulfate (DS) ions (from the SDS sample and running buffers) in the stacking gel. This zone of stacked DS micelles causes mixing of the DS ions with the smaller proteins, resulting in fuzzy bands and decreased resolution. The mixing also interferes with the fixing and staining of smaller proteins.

To help solve this problem, we offer the Novex® Tricine gel system that is based on the Tris-Glycine system developed by Schagger and von Jagow. This modified system uses a low pH in the gel buffer and substitutes tricine for glycine in the running buffer. The smaller proteins and peptides that migrate with the stacked DS micelles in the Tris-Glycine protein gel system are now well-separated from DS ions in the Novex® Tricine gel system, resulting in sharper bands and higher resolution.

The migration patterns of protein markers on Novex® Tricine, IEF, and zymogram gels are shown. Optimal resolution is achieved when protein bands migrate within the shaded regions. The numbered bands on the Tricine gel patterns correspond to the migration of Mark12™ Unstained Standard under denaturing conditions. The numbered bands on the zymogram gel patterns refer to the following proteases: Band 1: collagenase type I (140 kDa); Band 2: thermolysin (37 kDa); Band 3: chymotrypsin (30 kDa); Band 4: trypsin (19 kDa).

What is in the Novex® Tricine gel system?

The following reagents are used to perform electrophoresis with Novex® Tricine gels: