Tris-Glycine buffer 10× concentrate has been used as a transfer buffer for Western blotting.[1]
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. 8.3 Contains no methanol.
Proceedings of the National Academy of Sciences of the United States of America, 76(9), 4350-4354 (1979-09-01)
A method has been devised for the electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. The method results in quantitative transfer of ribosomal proteins from gels containing urea. For sodium dodecyl sulfate gels, the original band pattern was
Western blot protocol details protein transfer from gels to nitrocellulose, crucial for immunoblotting procedures in research.
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