Buffers Electrophoresis Buffer Systems Tris Glycine Tricine TBE Urea

TRIS-GLYCINE GELS

The Gel buffer is 0.375M Tris-HCl, pH 8.8 (Glycine is in the Running Buffer). Tris-Glycine Gels can be used for Denaturing or Non-Denaturing samples. Tris-Glycine Gels containing SDS can be purchased from Jule, Inc. ("D" in catalog number) or without SDS in the Gels ("W" in the catalog number).

Denaturing Gels ("D" in the part number, contains SDS): Gel buffer is 0.375M Tris-HCl, pH 8.8, 0.1 % SDS. Glycine is in the Running Buffer. These gels are used for the separation of proteins that have been exposed to SDS and therefore are in their denatured or unfolded state. These Gels enable the proteins to be separated on the basis of size alone. The proteins are no longer active, since they are unfolded. Add SDS to the sample and running buffer in order to run these gels as denaturing gels.

Non-Denaturing Gels ("W" in the part number): The Gel buffer is 0.375M Tris-HCl, pH 8.8 (No SDS in Gel Buffer).

Glycine is in the Running Buffer. Non-Denaturing Gels are used for separating proteins in their Native State. Native means any of the following: 1) The structure of a protein as it exists in nature; 2) the structure of a protein as isolated, if it retains enzymatic activity; 3) the form of a protein that has no biological activity but possesses secondary structure. These gels can be run as denaturing gels by treating the sample with a denaturing buffer and using running buffer that contains SDS. See proteins separated on Gels (Click here).

Tricine Gels:

("T" in the part number): Gel buffer is 1.0 M Tris-HCl, pH 8.45, 0.1 % SDS. These Gels are used for separating proteins or peptides as small as 1,000 Daltons (1kD). Tricine in the running buffer migrates faster than Glycine and the smallest proteins, thereby enabling separation. Use Tricine Gels for separating low molecular weight proteins (70,000 to 1,000 Daltons), but can be used for separating proteins from 212,000 to 1,000 Daltons depending upon the acrylamide gel concentration. Tricine Gels require using a different upper and lower running buffer (see Running Buffers). See proteins separated on Gels (Click here).

Non-Denaturing TBE Gels (for DNA, RNA, PCR product analysis, and proteins): ("N" in the part number) The Gel buffer is 0.090 Molar Tris, 0.080 Molar Boric Acid, pH 8.3, 0.0026M EDTA. These Gels are used for non-denaturing (native) protein analysis as described above under non-denaturing Tris-Glycine gels and non denaturing DNA and RNA analysis (double stranded). The applications for DNA include but are not limited to purification of DNA restriction fragments for cloning, PCR product analysis and purification, and gel mobility shift assays. These Gels use TBE Running Buffer.

Denaturing TBE/UREA Gels: Same as TBE Gels, except Urea (7 molar) is added. ("U" in the part number) These gels are used for single stranded DNA and RNA analysis. The Urea is a denaturing agent that causes the DNA and RNA to become single-stranded by breaking the bonds that usually hold the DNA and RNA together. The applications for this type of gel include: DNA size measurements and Southern Blot analysis, RNA size measurements and Northern Blot analysis, DNA foot printing (localizing of protein binding site on DNA or detection of DNA Binding proteins), RNA foot printing (location of site of RNA-binding proteins), and purification of oligonucleotides used for PCR and cDNA screening. These Gels use TBE Running Buffer.