SDS-PAGE, protein transfer, western Blot reagent preparation.

1.5 mol/L Tris (PH8.8)
1. Weighing 181.7g Tris is placed in
1L
begum.
2. Add about 800 ml of deionized water and stir to dissolve.
3. Adjust the pH to 8.8 with hydrochloric acid.
4. Set capacity to 1L.
5.
Hiculation
, save
at room temperature Note: the solution should be cooled to room temperature and then adjust the pH, because the pH of tris solution varies greatly with the temperature, the temperature increases by 1oC, the pH of the solution is reduced by about 0.03 units.
(see kara company Catalog-N1)
1 mol/L Tris (PH6.8)
1. Weigh 121.1g Tris in a 1L berries.
2. Add about 800 ml of deionized water and stir to dissolve.
3. Add the pH required for the adjustment of thick hydrochloric acid to the table below.
pH Thick HCl
6.8
7.4 about 70 ml
7.6 about 60 ml
8.0 about 42 ml
4. Set capacity to 1L.
5. After autobaric sterilization, store at room temperature.
note: 1. The pH of the solution is cooled to room temperature before the pH is adjusted, as the pH of the Tris solution varies greatly with the temperature, and for every 1oC increase in temperature, the pH of the solution decreases by approximately 0.03 units. 2. If the 1 mol/L solution is yellow, it should be discarded and equipped with better quality Tris.
(see kara company Catalog-N1)
30% acrylamide (100ml)
1. Weigh the following
reagents
in a beech.
acrylamide 29 g
biacrylamide (BIS) 1 g
2. Add about 60 ml of deionized water and stir to dissolve.
3. To 100 ml, filter out impurities with a 0.45 m membrane.
4. Store in a brown bottle of 4 oC.
note: 1. Acrylamide has a strong neurotoxicity, and can be absorbed through the skin, its effect is cumulative, preparation should wear
gloves
etc. Polyacrylamide is non-toxic, but should also be operated with caution as it is possible to contain small amounts of non-polymerized ingredients.
(refer to kara's Catalog-N1 and Molecular Clone II P924)
30%
SDS
1. SDS weighing 10 g high purity (electrophoresis grade) is placed in a 100 to 200 ml berries.
2. Add about 800 ml of deionized water, 68oC
heating
dissolve.
3. Add a few drops of thick hydrochloric acid to adjust the pH to 7.2.
4. Set the capacity to 100 ml and store at room temperature.
(refer to kara's Catalog-N1 and Molecular Clone II P924)
.