| Description | Medicago AB Products Are Available Worldwide. For Research Use Only. The most common running buffer in native (non-denaturing) homogeneous and gradient polyacrylamide gel electrophoresis (PAGE). Tris-Glycine also has applications in Western blotting. Tris-Glycine gels use a stacking gel to compress the sample into a narrow band before it enters the resolving gel. This leads to much sharper bands than would be seen in gels lacking a stacking gel. Tris-Glycine gels resolve proteins by charge/size. However, very small proteins and peptides do not resolve well due to interference from the glycine/pH discontinuity front. TG buffer is used to make a Tris-glycine/20% methanol Western transfer buffer, which is the most common protein transfer buffer for wet blot transfers. The methanol prevents the gel from swelling during transfer and enhances protein binding to nitrocellulose. Make sufficient transfer buffer to cover the electrode wires in the wet blot transfer unit and to soak the gel, membrane and blotting paper. Protein electrophoresis under denaturing conditions (SDS-PAGE) involves separating proteins based on their size. By treating the sample under denaturing and reducing conditions with sodium dodecyl sulfate (SDS), proteins unfold and become coated with SDS detergent molecules, thereby acquiring a high net negative charge that is proportional to the length of their polypeptide chain. During electrophoresis, the negatively-charged protein molecules migrate towards the positive electrode. |
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