Application

Tris-Glycine buffer 10× concentrate has been used as a transfer buffer for Western blotting.

Biochem/physiol Actions

For Western blotting and gel electrophoresis. Tris-glycine buffer is used to make a Tris-glycine-methanol transfer buffer, which is the most common protein transfer buffer for wet blot transfers. The methanol prevents the gel from swelling during the transfer and enhances the protein binding to nitrocellulose.The 10× Tris-glycine buffer is diluted to 1× with methanol and water to make a solution containing 25 mM Tris, 192 mM glycine, and 20% methanol. A sufficient amount of transfer buffer should be made to cover the electrode wires in the wet blot transfer unit and to soak the gel, membrane and blotting paper.

Other Notes

0.25 M Tris, 1.92 M glycine, pH approx. 8.3Contains no methanol.