Phosphate Buffered Saline (PBS) Preparation
Updated 6/12/2015 07:57 PM
Introduction
Materials
- Consumables
- sodium chloride (NaCl)
- potassium chloride (KCl)
- disodium phosphate (Na2HPO4)
- potassium phosphate monobasic (KH2PO4)
- calcuium chloride (CaCl2) (optional)
- magnesium chloride (MgCl2) (optional)
- Disposables
- weigh dishes/paper
- Equipment
- sterile container
- balance
- pH meter
Procedure
Components
Table1
A A | B B | |
1 1 | Final Volume (mL)* | 1000 |
2 2 | Final Stock Concentrate (x PBS)* | 10 |
Table2
A A | B B | C C | D D | |
1 1 | Component | Final Concentration (at 1xPBS) (mM)* | Hydrate(s)* | Amount (g) |
2 2 | sodium chloride | 137 | 80.0628 | |
3 3 | potassium chloride | 2.7 | 2.0128851 | |
4 4 | disodium phosphate | 10 | 0 | 14.196 |
5 5 | potassium phosphate monobasic | 2 | 2.72172 | |
6 6 | calcium chloride (optional) | 1 | 0 | 1.1098 |
7 7 | magnesium chloride (optional) | 0.5 | 0 | 0.476055 |
Note: It is possible to edit the desired concentrations, however it is not recommended for the non-optional components. These are the default concentrations for the traditional PBS solution.
Preparation
- Add ~80% of the final volume of dH2O to a sterile container.
Generally, the capacity of the container should be greater than (not equal to) the final volume of solution to be prepared in order to prevent the solution from boiling over during the autoclave step.
Ensure you have a volumetric flask measuring your desired final volume.
- Adjust the final volume and stock concentrate values in the table above and add the calculated amounts of the components to the dH2O.
- Titrate solution to pH 7.4 using concentrated HCl.
- Once completely dissolved, transfer the solution to a sterile volumetric flask and adjust the final volume using dH2O, then return it to the container.
Ensure the solution is well mixed before returning it to the container.
- Apply autoclave tape to the bottle and autoclave on liquids cycle.
- Store at room temperature.
This protocol describes the preparation of phosphate-buffered saline adapted from Sambrook & Rusell.
Note: Typical stock solution is 10x concentrated.
Sambrook, J. & Russell, D. W. (2001). Molecular Cloning: A Laboratory Manual, 3 edn. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press. pp. A1.7