Solutions

Buffered Salt Solution

10x PB, (500 ml)
(Final concentration for 1x is 10 mM NaH₂PO₄; 500 mM NaCl; pH 7.2)
Reference: Sigma Techniques for Product A-3665

Concentration of the 10x is 0.1 M Na-phosphate; 5 M NaCl.
400 ml dH₂O, set stirring and add:
6.9 g NaH₂PO₄ (fw 138 g/mol)
Make sure you use NaH₂PO₄ or you will need to recalculate!
146 g NaCl (add slowly)
pH 7.2 with 5 M NaOH at 1 ml at a time
Adjust volume to 500 ml in graduated cylinder

10x PBS (Phosphate-buffered Saline), (1 L)

80 g NaCl
2 g KCl
14.4 g Na₂HPO₄
2.4 g KH₂PO₄
Adjust to pH 7.2
Sterilize by autoclaving. Store at RT.

10x TBS (Tris-buffered Saline), (1 L)
(Final concentration for 1x is 25 mM Tris;137 mM NaCl; 3 mM KCl; pH 7.0)
Reference: Antibodies: A Laboratory Manual, page 684.

700 ml dH₂O, set stirring and add:
30.25 g Tris (fw 121 g/mol)
80.0 g NaCl (fw 58.4 g/mol)
2.24 g KCl (fw 74.6 g/mol)
Adjust to pH 7.0 with concentrated HCl (10 ml or perhaps more may be needed; CAREFUL)
To 1000 ml with dH₂O

For T-TBS, add Tween to 0.05%. This is most easily done by adding 1/100th volume from a stock solution of 5% tween (this can be pipetted easily). To make the 5% Tween, briefly warm the Tween 20 bottle in warm tap water. Cut the end off the pipet tip, and pipette slowly 750痞 of this stock into 14.25 ml TBS in a 15 ml tube.

5x ConA Buffer, (1 L)
(Final concentration for 1x is 20 mM Tris; 500 mM NaCl; 1 mM CaCl₂; 1 mM MgCl₂; pH 7.4)

700 ml dH₂O, set stirring and add:
100 ml 1 M Tris, pH 7.2-7.4
5 ml 1 M MgCl₂
5 ml 1 M CaCl₂
146 g NaCl
Check pH is between 7.2 and 7.4
To 1000 ml with dH₂O

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Last update Sept 27, 2000
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