Tricine Protein Gels

Invitrogen Novex Tricine Protein Gels provide separation of low molecular weight proteins and peptides. The Novex Tricine Gel System is a modification of the tris-glycine discontinuous buffer system developed by Schaegger and von Jagow (Schaegger and von Jagow, 1987), specifically designed for resolving peptides and low molecular weight proteins. In this system the tricine replaces the glycine in the running buffer, resulting in more efficient stacking and de-stacking of low molecular weight proteins and higher resolution of smaller peptides.

Features of Novex Tricine Protein Gels include:

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

How tricine gels work

In the traditional tris-glycine protein gel system, 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, which causes mixing of the DS ions with smaller proteins and results in fuzzy bands and decreased resolution. The mixing also interferes with the fixing and staining of smaller proteins. The Novex Tricine Gel 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 ions in the tris-glycine gel system are well separated from DS ions in the Novex Tricine Gel System, offering sharper bands and higher resolution.

Resolution of low molecular weight proteins using Tricine gels

The ability of Novex Tricine gels to resolve low molecular weight proteins was demonstrated in a comparison to Bio-Rad TGX 4-20% Tris-Glycine gels using samples of cleaved caspase-3. Samples were subjected to electrophoresis, followed by protein transfer and western blotting. Cleavage products of 19 kDa and 17 kDa resolved on the Novex 16%  Tricine gel while they did not on the Bio-Rad gel.

Novex Tricine gels better resolve low molecular weight proteins.

Lane 1: Thermo Scientific PageRuler™ Prestained NIR Protein Ladder;
Lanes 2-9: 1 μL loads per well of a 1.5x dilution series of a Jurkat cell lysate after cytochrome C treatment. After separation on Novex 16% Tricine and Bio-Rad TGX 4-20% Tris-Glycine gels, transfers were probed with primary antibodies labeled with Invitrogen Alexa Fluor™ Plus 680 and Invitrogen Alexa Fluor™ Plus 800 dyes, respectively. The Novex 16% Tricine gel resolved the 17 and 19 kDa bands of cleaved caspase-3, whereas the Bio-Rad TGX Tris-Glycine gel did not provide such resolution.