Dako Autostainer Plus - Manual (Page 105)

Dako Autostainer Plus | User Guide 105
Section 19 | Wash Buffers
Wash buffers are required for all immunohistochemical staining procedures. They function to eliminate
background by washing away the nonspecifically bound proteins from the specimen. Commonly used
buffers for immunohistochemical staining are Tris-buffered saline (TBS) or phosphate buffered saline
(PBS). These buffers are recommended by Dako for use on the Dako Autostainer Plus. The following
pages describe detailed instructions for preparation of TBS and PBS buffer solutions. Buffers used
with the Dako Autostainer Plus should contain a final concentration of 0.05% Tween 20.
Dako Autostainer Plus Wash Buffer
The Dako Autostainer Plus Wash Buffer is a Tris-buffered saline solution (TBS), pH 7.6, containing
0.05% Tween 20. It is recommended that Dako prepackaged buffer solutions are used for consistent
performance.
Intended Use
Dako Tris-Buffered Saline Solution containing 0.05% Tween 20 is used as the standard wash
buffer for the Dako Autostainer Plus.
Materials Required
1. Clean carboy, flask or beaker graduated to 1 Liter and 5 Liters
2. Deionized water
3. Magnetic stir plate with stir bar
4. Prepackaged Dako reagents
Code No. Contents
S3006 One Liter of Wash Buffer 10X. Each bottle makes 10 liters of 50mM Tris-HCI,
150mM NaCl, 0.05% Tris-Buffered Saline containing Tween 20 and a preservative
pH 7.6 or
S3001 Six packets of TBS containing Tris base, Tris-HCI and NaCl. Each packet makes
1 liter of 50mM Tris-HCI, 150mM NaCl Tris-Buffered Saline, pH 7.6 or
S1968 Two packets of TBS containing Tris base, Tris-HCI and NaCl. Each packet makes
1 liter of 50mM Tris-HCI, 150mM NaCl Tris-Buffered Saline, pH 1.6 or
S1966 Tween 20, 100mL.
Reagent Preparation
One Liter of Tris-Buffered Saline Solution (S3001)
1. Add 1L (1000 mL) of deionized water to a 1L 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 0.5 mL of Dako Tween 20 (Code No. S1966) to the Tris-buffered saline solution and
mix using a magnetic stir plate.