Dako Autostainer Plus - Manual (Page 106)

106 Dako Autostainer Plus | User Guide
4. The final solution is 50mM Tris-HCl, 150mM NaCl buffered saline, pH 7.6 with 0.05%
Tween 20.
Five Liters of Tris-Buffered Saline Solution (S1968)
1. Add 5 L (5000 mL) of deionized water to a 5 L carboy, flask or beaker.
2. Empty entire contents of one packet. Use a magnetic stir plate to mix and dissolve the
TBS granules.
3. Add 2.5 mL of Dako Tween 20 (Code No. S1966) to the Tris-buffered saline solution and
mix using a magnetic stir plate.
4. The final solution is 50 mM Tris-HCl, 150 mM NaCl buffered saline, pH 7.6 with 0.05%
Tween 20.
Ten Liters of Tris-Buffered Saline Solution (S3006)
1. Prior to dilution, allow the Wash Buffer 10X solution to reach room temperature (20-25°
C).
2. Add 1 L (1000 mL) of concentrated Wash Buffer 10X solution to a 10 L carboy, flask or
beaker.
3. Add 9 L (9000mL) of deionized water to the same container. Use a magnetic stir plate to
stir gently for at least 15 minutes.
4. The final solution is 50 mM Tris-HCl, 150 mM NaCl buffered saline, pH 7.6 with 0.05%
Tween and a preservative.
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 Plus 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 Plus.