Concentrated stock solution for tris/glycine/SDS gel run buffer (PAGE)

SDS denatures proteins and binds in a constant mass ratio to their peptide chains, covering their intrinsic charge. In the collection gel (pH 6.8) all proteins are kept in a sharp band between the neutral Glycin and chloride ions and thus all reach the separation gel at the same time. After the pH change when the separation gel is reached (pH 8.8), the chloride ions and the now cationic Glycin migrate very quickly to the anode and leave the proteins to themselves, which now move in the electric field depending on size towards the anode.

ROTIPHORESE^®10x SDS-PAGE contains 0,25 M Tris, 1,92 M glycine and 1 % (w/v) SDS in distilled, deionised water for protein analysis. Acc. to Laemmli (1970), Nature 227:680.

ROTIPHORESE^®10x SDS-PAGEis usually used as 1x concentrated solution. SDS-PAGE buffer is suitable for the denaturing separation of proteins and peptides of any size in the discontinuous polyacrylamide gel (collection gel / separation gel).