Preparation Phosphate Buffer Saline (PBS) 2013
Cell Culture Lab (VM3417) Procedure
Preparation of Phosphate Buffer Saline (PBS)
Investigator: __________
Date: ___________
Protocol: ______________________
1. A 1 liter stock of 10x PBS can be prepared by dissolving
- Add 80 g NaCl,
- Add 2 g KCl,
- Add 11.5 g Na2HPO4
- Add 2 g KH2PO4
- Add 1 Litter of Distilled water
After complete mixing, top up final solution to 1 L. The pH of the 10X stock is
will be approximately 6.8, but when diluted to 1x PBS it should change to 7.4. When
making buffer solutions, it is good practice to always measure the pH directly using a pH
meter. If necessary, pH can be adjusted using hydrochloric acid or sodium hydroxide.
Sterilize by autoclaving (20 min, 121°C). Store at room temperature.
2. For 1 liter of 1X PBS, prepare as follows:
Start with 800 ml of distilled water:
- Add 8 g of NaCl.
- Add 0.2 g of KCl.
- Add 1.15 g of Na2HPO4.
- Add 0.24 g of KH2PO4.
- Adjust the pH to 7.4 with HCl. And add distilled water to a total volume of 1
liter.
Dispense the solution into aliquots and sterilize by autoclaving (20 min, 121°C).
Store at room temperature.
Or used 10XPBS by add 100 ml of 10XPBS, and add 900 ml of Distilled water
sterilize by autoclaving (20 min, 121°C). Store at room temperature.
Notes:
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This procedure was revised on 24-06-2013 by Sunsanee Supunkong.