PBS: Difference between revisions

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(→‎Recipe: added dibasic heptahydrate)
 
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==Recipe==
==Recipe==
1x PBS is 137 mM NaCl, 10 mM Phosphate, 2.7 mM KCl, pH 7.4
1x PBS is 137 mM NaCl, 12 mM Phosphate, 2.7 mM KCl, pH 7.4


To make 10x PBS,
To make 10x PBS,
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#* 80 g NaCl
#* 80 g NaCl
#* 2 g KCl
#* 2 g KCl
#* 14.4 g Na<sub>2</sub>HPO<sub>4</sub>
#* 14.4 g Na<sub>2</sub>HPO<sub>4</sub> (dibasic anhydrous) OR 18.1 g Na<sub>2</sub>HPO<sub>4</sub> · 2H<sub>2</sub>O (dibasic dihydrate) OR 27.2 g Na<sub>2</sub>HPO<sub>4</sub> · 7H<sub>2</sub>O (dibasic heptahydrate)
#* 2.4 g KH<sub>2</sub>PO<sub>4</sub>
#* 2.4 g KH<sub>2</sub>PO<sub>4</sub> (monobasic anhydrous)
#* 800 mL distilled H<sub>2</sub>O
#* 800 mL distilled H<sub>2</sub>O
# Adjust pH to 7.4 with HCl
# Adjust pH to 7.4 with HCl
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*'''[[PBST]]''': For some applications (washing) one adds Tween 20 PBS which makes [[PBST]]  
*'''[[PBST]]''': For some applications (washing) one adds Tween 20 PBS which makes [[PBST]]  
**Usual concentrations of Tween 20 in TBST are 0.1 or 0.05%.
**Usual concentrations of Tween 20 in PBST are 0.1 or 0.05%.
 
**Usual concentration of Triton X-100 in TBST is 0.1-0.3%.
Another buffer used for washing in immunohistology is PBS Triton, which contains 0.1%-0.3% of the detergent Triton X-100. A 1% PBS Triton X-100 wash can also be added to fixed tissue sections for 10 minutes (followed by 3 time, 15-minute PBS wash) to permeabilize tissue prior to blocking at the beginning of a immunostaining experiment.


==Other Links==
==Other Links==
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* [http://en.wikipedia.org/wiki/Phosphate_buffered_saline wikipedia article]
* [http://en.wikipedia.org/wiki/Phosphate_buffered_saline wikipedia article]


[[Category:Material]] and [[Category:Buffers]]
[[Category:Material]] [[Category:Buffers]]

Latest revision as of 08:22, 22 April 2009

Purpose

PBS stands for phosphate buffered saline and is often used because it is isotonic to cells.

Recipe

1x PBS is 137 mM NaCl, 12 mM Phosphate, 2.7 mM KCl, pH 7.4

To make 10x PBS,

  1. Combine the following:
    • 80 g NaCl
    • 2 g KCl
    • 14.4 g Na2HPO4 (dibasic anhydrous) OR 18.1 g Na2HPO4 · 2H2O (dibasic dihydrate) OR 27.2 g Na2HPO4 · 7H2O (dibasic heptahydrate)
    • 2.4 g KH2PO4 (monobasic anhydrous)
    • 800 mL distilled H2O
  2. Adjust pH to 7.4 with HCl
  3. Add H2O to 1L
  4. Autoclave for 20 minutes on liquid cycle. Store at RT.


  • PBST: For some applications (washing) one adds Tween 20 PBS which makes PBST
    • Usual concentrations of Tween 20 in PBST are 0.1 or 0.05%.
    • Usual concentration of Triton X-100 in TBST is 0.1-0.3%.

Other Links

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