Phosphate buffered saline: Difference between revisions
add 2H2O at Na2HPO4 (if you take 144g, it's the dihydrate!), add referece pmid 13130792 |
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==Applications== | ==Applications== | ||
PBS has many uses because it is isotonic and non-toxic to cells. It can be used to dilute substances. It is used to rinse containers containing cells. PBS can be used as a diluent in methods to dry biomolecules, as water molecules within it will be structured around the substance (protein, for example) to be 'dried' and immobilized to a solid surface {{fact|date=April 2008}}. The thin film of water that binds to the substance prevents [[Denaturation_%28biochemistry%29 | denaturation]] or other conformational changes. Carbonate buffers may be used for the same purpose but with less effectiveness {{fact|date=April 2008}}. PBS can be used | PBS has many uses because it is isotonic and non-toxic to cells. It can be used to dilute substances. It is used to rinse containers containing cells. PBS can be used as a diluent in methods to dry biomolecules, as water molecules within it will be structured around the substance (protein, for example) to be 'dried' and immobilized to a solid surface {{fact|date=April 2008}}. The thin film of water that binds to the substance prevents [[Denaturation_%28biochemistry%29 | denaturation]] or other conformational changes. Carbonate buffers may be used for the same purpose but with less effectiveness {{fact|date=April 2008}}. PBS can be used to take a reference spectrum when measuring the protein adsorption in ellipsometry {{fact|date=April 2008}}. | ||
Additives can be used to add function. For example, PBS with [[EDTA]] is also used to disengage attached and clumped cells. Divalent [[metals]] such as [[zinc]], however, cannot be added as this will result in precipitation. For these types of applications, [[Good's buffers]] are recommended. | Additives can be used to add function. For example, PBS with [[EDTA]] is also used to disengage attached and clumped cells. Divalent [[metals]] such as [[zinc]], however, cannot be added as this will result in precipitation. For these types of applications, [[Good's buffers]] are recommended. | ||
Revision as of 09:45, 29 May 2008
Phosphate buffered saline (abbreviated PBS) is a buffer solution commonly used in biochemistry and other branches of biological research. It is a salty solution containing sodium chloride, sodium phosphate, and (in some formulations) potassium chloride and potassium phosphate. The buffer helps to maintain a constant pH. The osmolarity and ion concentrations of the solution usually match those of the human body (isotonic).
Applications
PBS has many uses because it is isotonic and non-toxic to cells. It can be used to dilute substances. It is used to rinse containers containing cells. PBS can be used as a diluent in methods to dry biomolecules, as water molecules within it will be structured around the substance (protein, for example) to be 'dried' and immobilized to a solid surface [citation needed]. The thin film of water that binds to the substance prevents denaturation or other conformational changes. Carbonate buffers may be used for the same purpose but with less effectiveness [citation needed]. PBS can be used to take a reference spectrum when measuring the protein adsorption in ellipsometry [citation needed].
Additives can be used to add function. For example, PBS with EDTA is also used to disengage attached and clumped cells. Divalent metals such as zinc, however, cannot be added as this will result in precipitation. For these types of applications, Good's buffers are recommended.
Preparation
There are many different ways to prepare PBS. Some formulations do not contain potassium, while others contain calcium or magnesium[1]. One of the most common preparations is described below.
A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na2HPO4 · 2H2O and 24 g KH2PO4 in 8 L of distilled water, and topping up to 10 L. The pH is ~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.
On dilution, the resultant 1x PBS should have a final concentration of 137 mM NaCl, 10 mM Phosphate, 2.7 mM KCl, and a pH of 7.4.
References
- ↑ Dulbecco, R. et al. (1954): Plaque formation and isolation of pure lines with poliomyelitis viruses. In: J. Exp. Med. vol. 99 (2), pp. 167-182. PMID 13130792
External links
pt:Tampão Fosfato-Salino de:PBS-Puffer fr:Tampon phosphate salin ja:リン酸緩衝生理食塩水