Tris-buffered saline (TBS) is appropriate for molecular biology since it is isotonic and non-toxic to cells. The buffer is frequently employed as a washing buffer or as a substance diluent in immunoassays. It is employed in immuno-histochemical identifying when the background is robust as well as for diluting alkaline phosphatase or peroxidase-conjugated antibodies in Western blotting.
Tris-buffered saline (TBS) is a buffer that is employed in several biochemical methods to keep the pH within a defined range. Tris has a mildly alkaline buffering capability in the 7-9.2 range (with HCl). At 25 °C, the conjugate acid of Tris has a pKa of 8.07. The pKa decreases by around 0.03 units every degree Celsius increase in temperature. When the temperature of the solution changes, the pH might alter significantly.
Applications of Tris Buffered Saline (TBS)
- Used as antibody diluent in Western blotting
- Used as a wash buffer in immunoassays
- Used in immuno-histochemical staining
- Used for sample diluent in in-vitro diagnostics
- Used in In situ hybridization
Recipe For the preparation of Tris Buffered Saline (TBS) 0.1 M
Reagents Required
| Ingredients | Amount | Concentration |
|---|---|---|
| Tris (mw: 121.14 g/mol) | 12.1 g | 0.09988 M |
| Sodium Chloride (NaCl) (mw: 58.44 g/mol) | 8 g | 0.1368 M |
| Potassium Chloride (mw: 74.55 g/mol) | 0.2 g | 0.002682 M |
Procedure For the Preparation of Tris Buffered Saline (TBS) 0.1 M
- 800 mL of distilled water should be ready in a convenient container.
- To the mixture, add 8 g of sodium chloride.
- 0.2 g of potassium chloride should be added to the mixture.
- Pour 12.1 g of Tris into the mixture.
- Adjust the pH to 7.4 with the use of HCl.
- Until the volume is 1 L, add distilled water.
Recipe For the Production of 10x Stock Tris Buffered Saline (TBS)
Reagents Required
| Ingredients | Amount | Concentration |
|---|---|---|
| Tris (mw: 121.14 g/mol) | 24 g | – |
| Sodium chloride (mw: 58.44 g/mol) | 88 g | – |
Procedure For the Production of 10x Stock Tris Buffered Saline (TBS)
- To prepare 1 L of 10X TBS stock solution, dissolve 24 g Tris and 88 g NaCl in 900 mL of distilled water.
- Secondly, adjust the pH of the solution to 7.6 with the help of HCl.
- Adjust the final volume to 1 L using the distilled water.
Video References
References
- https://www.sigmaaldrich.com/NP/en/support/calculators-and-apps/10x-tris-buffered-saline
- https://cshprotocols.cshlp.org/content/2009/6/pdb.rec11830.full
- https://www.sigmaaldrich.com/NP/en/support/calculators-and-apps/1x-tris-buffered-saline
- https://www.aatbio.com/resources/buffer-preparations-and-recipes/tris-buffered-saline-tbs-0-1-m
