Phosphate-buffered saline




Phosphate-buffered saline (abbreviated PBS) is a buffer solution commonly used in biological research. It is a water-based salt solution containing disodium hydrogen phosphate, sodium chloride and, in some formulations, potassium chloride and potassium dihydrogen phosphate. The buffer helps to maintain a constant pH. The osmolarity and ion concentrations of the solutions match those of the human body (isotonic).




Contents






  • 1 Applications


  • 2 Preparation


  • 3 See also


  • 4 References


  • 5 External links





Applications


PBS has many uses because it is isotonic and non-toxic to most cells. These uses include substance dilution and cell container rinsing. 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 solutions (one of them is Dulbecco's phosphate-buffered saline (DPBS), which has a lower phosphate concentration than standard PBS[1]). Some formulations do not contain potassium and magnesium, while other ones contain calcium and/or magnesium.































The most common composition of PBS (1X)
Salt Concentration (mmol/L) Concentration (g/L)
  NaCl   137 8.0
  KCl   2.7 0.2
  Na2HPO4   10 1.44
  KH2PO4   1.8 0.24

Start with 800 mL of distilled water to dissolve all salts. Add distilled water to a total volume of 1 liter. The resultant 1x PBS will have a final concentration of 157 mM Na+, 140mM Cl−, 4.45mM K+, 10.1 mM HPO42−, 1.76 mM H2PO4− and a pH of 7.96. Add 2.84mM of NaCl to shift the buffer to 7.3 mM HPO42− and 4.6 mM H2PO4− for a final pH of 7.4 and a Cl− concentration of 142 mM.

























































Cold Spring Harbor Protocol[2]
reagent MW mass (g) 10X [M] 10X mass (g) 5X [M] 5X mass (g) 1X [M] 1X
Na2HPO4
 141.95897 14.1960 0.1000 7.0980 0.0500 1.41960 0.0100
KH2PO4
 136.08569 2.4496 0.0180 1.2248 0.0090 0.24496 0.0018
NaCl 58.44300 80.0669 1.3700 40.0335 0.6850 8.00669 0.1370
KCl 74.55150 2.0129 0.0270 1.0064 0.0135 0.20129 0.0027
pH = 7.4

The pH of PBS is ~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.


PBS can also be prepared by using commercially made PBS buffer tablets or pouches.[3]


If used in cell culturing, the solution can be dispensed into aliquots and sterilized by autoclaving or filtration. Sterilization may not be necessary depending on its use. PBS can be stored at room temperature or in the refrigerator. However, concentrated stock solutions may precipitate when cooled and should be kept at room temperature until precipitate has completely dissolved before use.



See also



  • Borate-buffered saline

  • Tris-buffered saline



References





  1. ^ Dulbecco, R.; et al. (1954). "Plaque formation and isolation of pure lines with poliomyelitis viruses". J. Exp. Med. 99 (2): 167–182. doi:10.1084/jem.99.2.167. PMC 2180341. PMID 13130792..mw-parser-output cite.citation{font-style:inherit}.mw-parser-output .citation q{quotes:"""""""'""'"}.mw-parser-output .citation .cs1-lock-free a{background:url("//upload.wikimedia.org/wikipedia/commons/thumb/6/65/Lock-green.svg/9px-Lock-green.svg.png")no-repeat;background-position:right .1em center}.mw-parser-output .citation .cs1-lock-limited a,.mw-parser-output .citation .cs1-lock-registration a{background:url("//upload.wikimedia.org/wikipedia/commons/thumb/d/d6/Lock-gray-alt-2.svg/9px-Lock-gray-alt-2.svg.png")no-repeat;background-position:right .1em center}.mw-parser-output .citation .cs1-lock-subscription a{background:url("//upload.wikimedia.org/wikipedia/commons/thumb/a/aa/Lock-red-alt-2.svg/9px-Lock-red-alt-2.svg.png")no-repeat;background-position:right .1em center}.mw-parser-output .cs1-subscription,.mw-parser-output .cs1-registration{color:#555}.mw-parser-output .cs1-subscription span,.mw-parser-output .cs1-registration span{border-bottom:1px dotted;cursor:help}.mw-parser-output .cs1-ws-icon a{background:url("//upload.wikimedia.org/wikipedia/commons/thumb/4/4c/Wikisource-logo.svg/12px-Wikisource-logo.svg.png")no-repeat;background-position:right .1em center}.mw-parser-output code.cs1-code{color:inherit;background:inherit;border:inherit;padding:inherit}.mw-parser-output .cs1-hidden-error{display:none;font-size:100%}.mw-parser-output .cs1-visible-error{font-size:100%}.mw-parser-output .cs1-maint{display:none;color:#33aa33;margin-left:0.3em}.mw-parser-output .cs1-subscription,.mw-parser-output .cs1-registration,.mw-parser-output .cs1-format{font-size:95%}.mw-parser-output .cs1-kern-left,.mw-parser-output .cs1-kern-wl-left{padding-left:0.2em}.mw-parser-output .cs1-kern-right,.mw-parser-output .cs1-kern-wl-right{padding-right:0.2em}


  2. ^ CSH Protocol: Phosphate-buffered saline (PBS) recipe


  3. ^ Medicago AB, (2010) Phosphate buffered saline specification sheet



  • Cold Spring Harbor Protocols


External links


  • http://www.bioind.com/products/cell-culture/cell-culture-reagents/balanced-salt-solutions/dpbs-dulbecco-s-phosphate-buffered-saline/



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