Dako Autostainer - Manual (Page 106)

106 Dako Autostainer | User Guide
6. The final solution is 0.5M Tris-HCl, 1.5M NaCl, buffered saline, pH 7.6 with
0.5% Tween 20.
One Liter of Tris-Buffered Saline, containing 0.05% Tween 20 from 10x stock
1. Add 900 mL of deionized water to a 1L (1000 mL) carboy, flask or beaker.
2. Add 100 mL of 10X Tris-buffered saline solution and mix using a magnetic stir plate.
3. The final solution is 50mM Tris-HCl, 150mM NaCl buffered saline, pH 7.6 with 0.05%
Tween 20.
One Liter Tris-Buffered Saline containing 0.05% Tween 20
1. Add approximately 500 mL of deionized water to a 1L (1000 mL) carboy, flask or beaker.
2. Dissolve 1.39 g of Trizma base, 6.06 g of Trizma HCl and 8.77 g of NaCl and dissolve
using a magnetic stir plate.
3. After all granules are dissolved adjust the solution to pH 7.6 with 12N HCl. Allow the
solution to continue to mix on the stir plate. Add the 12N HCl dropwise, observing the pH
constantly.
4. Once pH 7.6 is reached, add 0.5 mL of Tween 20 to the Tris-buffered saline solution and
mix using a magnetic stir plate.
5. Adjust the final volume of buffer to 1 liter with deionized water.
6. The final solution is 50mM Tris-HCl, 150mM NaC, buffered saline, pH 7.6 with 0.05%
Tween 20.
Shelf Life and Storage
TBS solutions should be stored at 2-8 °C to inhibit bacterial growth. TBS solutions that become
turbid should be discarded.
If Tris-buffered saline is stored at 4
°C, the expected shelf life is 7 days.
If Tris-buffered saline is stored at room temperature, the expected shelf life is 4 days.
NOTE: The above formulation for Tris-buffered saline does not contain any preservatives. It is
recommended that the wash buffer carboy is washed according to good laboratory practices each
time before it is filled with fresh buffer. Also, it is recommended that the end piece of the in-line
buffer tubing is wiped clean periodically to avoid clogging.