Dako Autostainer Plus - Manual (Page 107)

Dako Autostainer Plus | User Guide 107
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 HCI Sigma, T-3252
NaCl Sigma, T-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 1 L (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.
6. The final solution is 0.5 M Tris-HCl, 1.5 M 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
7. Add 900 mL of deionized water to a 1 L (1000 mL) carboy, flask or beaker.
8. Add 100 mL of 10X Tris-buffered saline solution and mix using a magnetic stir plate.
9. The final solution is 50 mM Tris-HCl, 150 mM 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 1 L (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 12 N HCl dropwise, observing