Western Blot Buffers
SDS-PAGE running buffer
To prepare 1 L of SDS-PAGE SDS Running Buffer (5x):
Reagent | Weight | Concentration |
---|---|---|
Tris | 30.2 g | 0.25 M |
Glycine | 188.0 g | 2.5 M |
SDS | 10 g | 10% |
Tris (tris(hydroxymethyl)aminomethane): C4H11NO3, 121.136 g/mol
Glycine: C2H5NO2, 75.067 g/mol
SDS (Sodium dodecyl sulfate): C12H25NaSO4, 288.372 g/mol
For 1x SDS-PAGE running buffer:
- Mix 200 mL 5x buffer with 800 mL water.
Tris-Glycine Transfer buffer (10x)
To prepare 1 L of Transfer buffer (10x)
Reagent | Weight | Concentration |
---|---|---|
Tris | 58 g | 480 mM |
Glycine | 29 g | 390 mM |
SDS | 3.7 g | 0.37 % |
- Mix 100 mL 10x buffer, 200 mL methanol and 700 mL cold water. (It works best for semi-dry transfer)
TBS buffer
To prepare 1 L of TBS Buffer (10x):
Reagent | Weight | Concentration |
---|---|---|
Tris | 24 g | 200 mM |
NaCl | 88 g | 1500 mM |
Tris (tris(hydroxymethyl)aminomethane): C4H11NO3, 121.136 g/mol
Glycine: C2H5NO2, 75.067 g/mol
- Adjust pH to 7.6 with HCl.
For 1x TBS running buffer:
- Mix 100 mL 10x buffer with 900 mL water.
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