Tris-buffered saline (TBS) is a widely used buffer in molecular biology, immunology, and biochemistry. It serves as a stable isotonic solution suitable for applications such as Western blotting, immunohistochemistry, and ELISA. The 10X TBS stock solution is a concentrated formulation that allows for efficient storage and easy dilution into working concentrations. The 10X TBS recipe calculator facilitates accurate preparation of the buffer by automating component calculations based on desired final volumes and molarities.
Desired Volume
Composition of 10X TBS Solution
A standard 10X TBS buffer typically consists of the following components:
- Tris Base (Tris(hydroxymethyl)aminomethane): Acts as a buffering agent to maintain a stable pH.
- Sodium Chloride (NaCl): Provides ionic strength to mimic physiological conditions.
- Hydrochloric Acid (HCl): Used for pH adjustment.
- Deionized Water (H2O): Serves as the solvent.
| Component | Concentration (mM) | Amount per 1 L |
| Tris Base | 200 | 24.2 g |
| NaCl (ACM7647145) | 1500 | 87.6 g |
| HCl | To adjust pH | Variable |
| H2O (ACM7732185) | - | To 1 L total |
Preparation Method for 10X TBS Solution
- Dissolve Tris Base: Weigh a quantity of Tris Base (can be calculated using the calculator) and add it to a beaker of an appropriate amount of deionized water.
- Dissolve NaCl: Add a quantity of NaCl to the same solution (can be calculated using the calculator) and stir until completely dissolved.
- Adjust pH: Using a calibrated pH meter, slowly add concentrated HCl while stirring until pH reaches 7.6.
- Volumizing: Transfer the solution to a calibrated vessel, add deionized water, and adjust the total volume to the desired target volume.
- Sterilization (optional): If sterilization is required, filter the solution through a 0.22 μm filter.
- Storage: Store 10X TBS solution at room temperature or 4°C for up to 3 months.
Adjusting the pH
Since Tris buffer has pH-dependent properties, HCl is added carefully while monitoring pH using a calibrated pH meter. The pH of a Tris buffer solution varies with temperature, so adjustments should be made at the working temperature.
Temperature Considerations:
| Temperature (°C) | Tris pKa |
| 4°C | 8.4 |
| 25°C | 8.1 |
| 37°C | 7.9 |
Applications of TBS in Research
- Western Blotting: Used as a washing buffer in immunodetection protocols.
- ELISA: Provides an isotonic environment for antibody-antigen interactions.
- Immunohistochemistry: Maintains physiological conditions for tissue staining.
- Cell Culture Rinsing: Removes unwanted residues while maintaining osmolarity.
