Different Buffer preparation for Gel Electrophoresis

Acrylamide Stock Solution Preparation

Application: Acrylamide is used along with bisacrylamide to form gels used in SDS-PAGE. Acrylamide forms polyacrylamide polymers, while bisacrylamide crosslinks between polyacrylamide chains.

To prepare 1L of Acrylamide Stock Solution the following components are required;

Acrylamide Stock Solution Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 389.6 g of acrylamide to the solution.
3. Add 10.4 g of bisacrylamide to the solution.
4. Add distilled water until volume is 1 L.
5. Filter the stock solution using Whatman filter paper and store at 4°C. Prepare fresh acrylamide stock solution every few weeks. T denotes the total percentage concentration of the monomers (acrylamide plus N,N’-methylenebisacrylamide) in grams per 100 mL. C denotes the percentage (by weight) of the cross-linker N,N’-methylenebisacrylamide relative to the total amount of monomers.

Electrode Buffer Preparation

Application: Electrode buffer has been used in studies involving polymer solar cells.

To prepare 1L of Electrode Buffer the following components are required;

Electrode Buffer Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 11.6 g of Tris base to the solution.
3. Add 37.5 g of glycine to the solution.
4. Add 3.98 g of SDS to the solution.
5. Add distilled water until volume is 1 L.

Electrophoretic Transfer Buffer (20x) Preparation 

Application: Electrophoretic Transfer Buffers are used in SDS-PAGE to transfer proteins from a gel to a nitrocellulose membrane.

To prepare 1L of Electrophoretic Transfer Buffer (20x) the following components are required;

Electrophoretic Transfer Buffer (20x) Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 35.29 g of Trisodium citrate to the solution.
3. Add 22.71 g of Sodium phosphate, dibasic (Na2HPO4) to the solution.
4. Add distilled water until volume is 1 L.
5. Adjust the pH to 8.3, autoclave, and store for up to 1-2 yr at room temperature.

Imidazole-SDS Solution Preparation

 To prepare 1L of Imidazole-SDS Solution the following components are required;

Imidazole-SDS Solution Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 13.6 g of imidazole to the solution.
3. Add 1 g of SDS to the solution.
4. Add distilled water until volume is 1 L.

SDS-PAGE SDS Running Buffer (10x) Preparation

Application: In SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), SDS Running Buffer is used as the electrophoresis buffer during stacking and resolution.

 To prepare 1L of SDS-PAGE SDS Running Buffer (10x) the following components are required;

SDS-PAGE SDS Running Buffer (10x) Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 30.3 g of Tris base to the solution.
3. Add 144.4 g of Glycine to the solution.
4. Add 10 g of SDS to the solution.
5. Add distilled water until volume is 1 L.

Sodium Carbonate Transfer Buffer (40x, pH 9.5) Preparation

Application: Sodium Carbonate Transfer Buffer is used to transfer proteins following SDS-PAGE.

To prepare 1L of Sodium Carbonate Transfer Buffer (40x, pH 9.5) the following components are required;

Sodium Carbonate Transfer Buffer (40x, pH 9.5) Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 21.1 g of NaHCO3 to the solution.
3. Add 18.35 g of Na2CO3 to the solution.
4. Adjust the pH to 9.5 with 10 N NaOH.
5. Add distilled water until volume is 1 L.
6. Store at room temperature.

Tris-Glycine Transfer Buffer (20x) Preparation 

Application: Tris-Glycine Transfer Buffer is used as a transfer buffer in Western blotting.

To prepare 1L of Tris-Glycine Transfer Buffer (20x) the following components are required;

Tris-Glycine Transfer Buffer (20x) Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 24.2 g of Tris base to the solution.
3. Add 150.1 g of Glycine to the solution.
4. Add distilled water until volume is 1 L.
5. pH adjustment is not necessary (it will be ~8.8). Store at room temperature.

Tris/EACA Buffer Preparation

To prepare 1L of Tris/EACA Buffer Preparation the following components are required;

Tris/EACA Buffer Preparation Steps

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1.31 g of ε-aminocaproic acid (EACA) to the solution.
3. Add 10.9 g of Tris base to the solution.
4. Add distilled water until volume is 1 L.