SDS-PAGE running buffer contains Tris Base, Glycine, and sodium dodecyl sulfate (SDS). It is used to run protein in SDS-PAGE in denaturing conditions. A 10x concentrated stock of SDS-PAGE running buffer is prepared by dissolving 30.3 g of Tris base, 144.0 g Glycine, and 10.0 g of SDS in water to a final volume of 1L. 1x SDS-PAGE running buffer is prepared by diluting the 10x stock solution.
Reagents and solutions
Tris base (Molecular Weight: 121.14) (e.g., Sigma-Aldrich #T1503)
Glycine (Molecular Weight: 75.07) (e.g., Sigma-Aldrich #G8898)
Sodium dodecyl sulfate (SDS) (Molecular Weight: 288.38) (e.g., Sigma-Aldrich #L3771)
Deionized / Distilled water
* Tris base (CAS Number: 77-86-1) and Trizma® base (CAS Number: 77-86-1) are the same chemicals.
Equipment and disposables
Magnetic stirrer with a heating device
0.2501 M Tris base
1.924 M Glycine
0.03467 M Sodium dodecyl sulfate (SDS)
Preparation of 1L of 10x SDS-PAGE Running Buffer
Step 1: Weigh out 30.3 g of Tris base, 144.0 g Glycine, and 10.0 g of SDS in a 2 L beaker. Add 750 ml Deionized/Distilled water and mix using a magnetic stirrer until all components dissolve completely.
Step 2: Adjust the final volume to 1L with Deionized/Distilled water. Mix it again. The pH of the solution should be 8.3 without any pH adjustment.