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;
Component | Amount | Concentration |
acrylamide (mw: 71.08 g/mol) | 389.6 g | 5.481 M |
bisacrylamide (mw: 154.17 g/mol) | 10.4 g | 0.0675 M |
Acrylamide Stock Solution Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 389.6 g of acrylamide to the solution.
- Add 10.4 g of bisacrylamide to the solution.
- Add distilled water until volume is 1 L.
- 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;
Component | Amount | Concentration |
Tris base (mw: 121.14 g/mol) | 11.6 g | 0.096 M |
glycine (mw: 75.07 g/mol) | 37.5 g | 0.5 M |
SDS (mw: 288.38 g/mol) | 3.98 g | 0.0138 M |
Electrode Buffer Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 11.6 g of Tris base to the solution.
- Add 37.5 g of glycine to the solution.
- Add 3.98 g of SDS to the solution.
- 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;
Component | Amount | Concentration |
Trisodium citrate (mw: 258.07 g/mol) | 35.29 g | 0.12 M |
Sodium phosphate, dibasic (Na2HPO4) (mw: 141.96 g/mol) | 22.71 g | 0.16 M |
Electrophoretic Transfer Buffer (20x) Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 35.29 g of Trisodium citrate to the solution.
- Add 22.71 g of Sodium phosphate, dibasic (Na2HPO4) to the solution.
- Add distilled water until volume is 1 L.
- 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;
Component | Amount | Concentration |
imidazole (mw: 68.08 g/mol) | 13.6 g | 0.2 M |
SDS (mw: 288.38 g/mol) | 1 g | 0.0035 M |
Imidazole-SDS Solution Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 13.6 g of imidazole to the solution.
- Add 1 g of SDS to the solution.
- 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;
Component | Amount | Concentration |
Tris base (mw: 121.14 g/mol) | 30.3 g | 0.2501 M |
Glycine (mw: 75.07 g/mol) | 144.4 g | 1.924 M |
SDS (mw: 288.38 g/mol) | 10 g | 0.0347 M |
SDS-PAGE SDS Running Buffer (10x) Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 30.3 g of Tris base to the solution.
- Add 144.4 g of Glycine to the solution.
- Add 10 g of SDS to the solution.
- 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;
Component | Amount | Concentration |
NaHCO3 (mw: 84.01 g/mol) | 21.1 g | 0.251 M |
Na2CO3 (mw: 105.99 g/mol) | 18.35 g | 0.173 M |
Sodium Carbonate Transfer Buffer (40x, pH 9.5) Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 21.1 g of NaHCO3 to the solution.
- Add 18.35 g of Na2CO3 to the solution.
- Adjust the pH to 9.5 with 10 N NaOH.
- Add distilled water until volume is 1 L.
- 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;
Component | Amount | Concentration |
Tris base (mw: 121.14 g/mol) | 24.2 g | 0.2 M |
Glycine (mw: 75.07 g/mol) | 150.1 g | 2 M |
Tris-Glycine Transfer Buffer (20x) Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 24.2 g of Tris base to the solution.
- Add 150.1 g of Glycine to the solution.
- Add distilled water until volume is 1 L.
- 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;
Component | Amount | Concentration |
ε-aminocaproic acid (EACA) (mw: 131.17 g/mol) | 1.31 g | 0.01 M |
Tris base (mw: 121.14 g/mol) | 10.9 g | 0.09 M |
Tris/EACA Buffer Preparation Steps
- Prepare 800 mL of distilled water in a suitable container.
- Add 1.31 g of ε-aminocaproic acid (EACA) to the solution.
- Add 10.9 g of Tris base to the solution.
- Add distilled water until volume is 1 L.