Phosphate buffered saline - Revision history

wikademia>Eme: /* Preparation */

2014-11-23T10:35:52Z

Preparation

← Older revision		Revision as of 10:35, 23 November 2014
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	==Preparation==		==Preparation==
	There are many different ways to prepare PBS. Some formulations do not contain potassium, while others contain calcium or magnesium<ref>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</ref>. One of the most common preparations is described below.		There are many different ways to prepare PBS. Some formulations do not contain potassium, while others contain calcium or magnesium<ref>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</ref>. One of the most common preparations is described below.


	The simplest way to prepare a PBS solution is to use PBS buffer tablets. They are formulated to give a ready to use PBS solution upon dissolution in a specified quantity of distilled water. They are available in the standard volumes: 100, 200, 500 and 1000 ml <ref>[http://www.medicago.se/_filer/PBS28-eng.pdf Medicago AB, (2003) Phosphate buffered saline pH 7.4 specification sheet]</ref>.		The simplest way to prepare a PBS solution is to use PBS buffer tablets. They are formulated to give a ready to use PBS solution upon dissolution in a specified quantity of distilled water. They are available in the standard volumes: 100, 200, 500 and 1000 ml <ref>[http://www.medicago.se/_filer/PBS28-eng.pdf Medicago AB, (2003) Phosphate buffered saline pH 7.4 specification sheet]</ref>.

wikademia>Eme at 10:35, 23 November 2014

2014-11-23T10:35:34Z

← Older revision		Revision as of 10:35, 23 November 2014
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	The simplest way to prepare a PBS solution is to use PBS buffer tablets. They are formulated to give a ready to use PBS solution upon dissolution in a specified quantity of distilled water. They are available in the standard volumes: 100, 200, 500 and 1000 ml <ref>[http://www.medicago.se/_filer/PBS28-eng.pdf Medicago AB, (2003) Phosphate buffered saline pH 7.4 specification sheet]</ref>.		The simplest way to prepare a PBS solution is to use PBS buffer tablets. They are formulated to give a ready to use PBS solution upon dissolution in a specified quantity of distilled water. They are available in the standard volumes: 100, 200, 500 and 1000 ml <ref>[http://www.medicago.se/_filer/PBS28-eng.pdf Medicago AB, (2003) Phosphate buffered saline pH 7.4 specification sheet]</ref>.


	A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na<sub>2</sub>HPO<sub>4</sub> and 24 g KH<sub>2</sub>PO<sub>4</sub> 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]].		A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na<sub>2</sub>HPO<sub>4</sub> and 24 g KH<sub>2</sub>PO<sub>4</sub> 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, 2.7 mM KCl, 10 mM Sodium Phosphate dibasic, 2 mM Potassium Phosphate monobasic and a pH of 7.4.		On dilution, the resultant 1x PBS should have a final concentration of 137 mM NaCl, 2.7 mM KCl, 10 mM Sodium Phosphate dibasic, 2 mM Potassium Phosphate monobasic and a pH of 7.4.


	Another preparation is described in Molecular Cloning by Sambrook, Fritsch and Maniatis, Apendix B.12<ref>Sambrook, Fritsch, and Maniatis (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, volume 3, appendix B.12</ref> as follows:		Another preparation is described in Molecular Cloning by Sambrook, Fritsch and Maniatis, Apendix B.12<ref>Sambrook, Fritsch, and Maniatis (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, volume 3, appendix B.12</ref> as follows:

	For 1 litre of Phosphate-buffered saline (PBS buffer) use:..		For 1 litre of Phosphate-buffered saline (PBS buffer) use:..
	- Dissolve in 800 ml of distilled H<sub>2</sub>O:		*Dissolve in 800 ml of distilled H<sub>2</sub>O:
	- 8 g of NaCl		*8 g of NaCl
	- 0.2 g of KCl		*0.2 g of KCl
	- 1.44 g of Na<sub>2</sub>HPO<sub>4</sub>		*1.44 g of Na<sub>2</sub>HPO<sub>4</sub>
	- 0.24 g of KH<sub>2</sub>PO<sub>4</sub>		*0.24 g of KH<sub>2</sub>PO<sub>4</sub>
	- Adjust the pH to 7.4 with HCl or NaOH		*Adjust the pH to 7.4 with HCl or NaOH
	- Add H<sub>2</sub>O to 1 liter.		*Add H<sub>2</sub>O to 1 liter.

	Dispense the solution into aliquots and sterilize them by autoclaving (20 min, 121°C, liquid cycle).		Dispense the solution into aliquots and sterilize them by autoclaving (20 min, 121°C, liquid cycle).
	Store at room temperature.		Store at room temperature.


	[[Category:Buffers]]		[[Category:Buffers]]

wikademia>Wikademia at 14:08, 26 April 2009

2009-04-26T14:08:07Z

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 Dispense the solution into aliquots and sterilize them by autoclaving (20 min, 121°C, liquid cycle).
 Store at room temperature.
 [[Category:Buffers]]

wikademia>Wikademia at 15:53, 24 April 2009

2009-04-24T15:53:45Z

← Older revision		Revision as of 15:53, 24 April 2009
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	'''Phosphate buffered saline''' (abbreviated PBS) is a [[buffer solution]] commonly used in [[biology \| 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]]).		'''Phosphate buffered saline''' (abbreviated PBS) is a [[buffer solution]] commonly used in [[biology \| 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]]).
			{{ntnes}}
	==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 to take a reference spectrum when measuring the protein adsorption in ellipsometry {{fact\|date=April 2008}}.		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}}.

wikademia>Wikademia: 104 revisions

2009-04-18T15:55:23Z

104 revisions

← Older revision	Revision as of 15:55, 18 April 2009
(No difference)

84.36.212.5: /* Preparation */

2009-04-05T10:18:05Z

Preparation

← Older revision		Revision as of 10:18, 5 April 2009
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	Another preparation is described in Molecular Cloning by Sambrook, Fritsch and Maniatis, Apendix B.12<ref>Sambrook, Fritsch, and Maniatis (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, volume 3, appendix B.12</ref> as follows:		Another preparation is described in Molecular Cloning by Sambrook, Fritsch and Maniatis, Apendix B.12<ref>Sambrook, Fritsch, and Maniatis (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, volume 3, appendix B.12</ref> as follows:

	For 1 litre of Phosphate-buffered saline (PBS buffer) use:		For 1 litre of Phosphate-buffered saline (PBS buffer) use:..
	- Dissolve in 800 ml of distilled H<sub>2</sub>O:		- Dissolve in 800 ml of distilled H<sub>2</sub>O:
	- 8 g of NaCl		- 8 g of NaCl

140.247.27.105: /* Preparation */

2009-04-02T16:50:53Z

Preparation

← Older revision		Revision as of 16:50, 2 April 2009
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	- 1.44 g of Na<sub>2</sub>HPO<sub>4</sub>		- 1.44 g of Na<sub>2</sub>HPO<sub>4</sub>
	- 0.24 g of KH<sub>2</sub>PO<sub>4</sub>		- 0.24 g of KH<sub>2</sub>PO<sub>4</sub>
	- Adjust the pH to 7.4 with HCl		- Adjust the pH to 7.4 with HCl or NaOH
	- Add H<sub>2</sub>O to 1 liter.		- Add H<sub>2</sub>O to 1 liter.

140.247.27.105: /* Preparation */ Correction of Na Phosphate type, which should be anhydrous, not 12 H2O for the mass in the recipes.

2009-04-02T16:28:01Z

Preparation: Correction of Na Phosphate type, which should be anhydrous, not 12 H2O for the mass in the recipes.

← Older revision		Revision as of 16:28, 2 April 2009
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	A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na<sub>2</sub>HPO<sub>4</sub>~~ · 12H<sub>2</sub>O~~ and 24 g KH<sub>2</sub>PO<sub>4</sub> 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]].		A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na<sub>2</sub>HPO<sub>4</sub> and 24 g KH<sub>2</sub>PO<sub>4</sub> 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, 2.7 mM KCl, 10 mM Sodium Phosphate dibasic, 2 mM Potassium Phosphate monobasic and a pH of 7.4.		On dilution, the resultant 1x PBS should have a final concentration of 137 mM NaCl, 2.7 mM KCl, 10 mM Sodium Phosphate dibasic, 2 mM Potassium Phosphate monobasic and a pH of 7.4.

140.247.27.105: /* Preparation */ Correction of concentrations to match Sambrook et al.

2009-04-02T16:25:56Z

Preparation: Correction of concentrations to match Sambrook et al.

← Older revision		Revision as of 16:25, 2 April 2009
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	A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na<sub>2</sub>HPO<sub>4</sub> · 12H<sub>2</sub>O and 24 g KH<sub>2</sub>PO<sub>4</sub> 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]].		A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na<sub>2</sub>HPO<sub>4</sub> · 12H<sub>2</sub>O and 24 g KH<sub>2</sub>PO<sub>4</sub> 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.		On dilution, the resultant 1x PBS should have a final concentration of 137 mM NaCl, 2.7 mM KCl, 10 mM Sodium Phosphate dibasic, 2 mM Potassium Phosphate monobasic and a pH of 7.4.

140.247.27.105: /* Preparation */

2009-04-02T16:22:16Z

Preparation

← Older revision		Revision as of 16:22, 2 April 2009
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	- 8 g of NaCl		- 8 g of NaCl
	- 0.2 g of KCl		- 0.2 g of KCl
	- 1.44 g of Na<sub>2</sub>HPO<sub>4</sub>~~and~~		- 1.44 g of Na<sub>2</sub>HPO<sub>4</sub>
	- 0.24 g of KH<sub>2</sub>PO<sub>4</sub>		- 0.24 g of KH<sub>2</sub>PO<sub>4</sub>
	- Adjust the pH to 7.4 with HCl		- Adjust the pH to 7.4 with HCl