Dako Autostainer - Manual (Page 105)

Dako Autostainer | User Guide 105
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: Dako Autostainer Wash Buffer 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.
Tris-Buffered Saline
The following is a procedure for the formulation of 50mM Tris, 150mM NaCl, buffered saline,
pH 7.6 with 0.05% Tween 20 from common laboratory chemicals.
Intended Use
Dako recommends a Tris-buffered saline solution (TBS), pH 7.6, containing 0.05% Tween 20 as
wash buffer for IHC procedures performed on the Autostainer.
Materials Required
1. Clean carboy, flask or beaker graduated to 1L
2. Deionized water
3. Magnetic stir plate with stir bar
4. Common laboratory chemicals
Chemical Manufacturer and Product No.
Trizma Base Sigma, T-1503
Trizma HCl Sigma, T-3253
NaCl Sigma, S-9625
Tween 20 Dako, S1966
Hydrochloric Acid 12N EM, HX0603-3
Reagent Preparation
One Liter of 10x Tris-Buffered Saline containing 0.5% Tween 20
1. Add approximately 500 mL of deionized water to a 1L (1000 mL) carboy, flask or beaker.
2. Add 13.9 g of Trizma base, 60.6 g of Trizma HCl and 87.66 g of NaCl and dissolve using
a magnetic stir plate.
3. After all granules are dissolved adjust the solution to pH 7.6 with 12 N 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 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.