Buffer Recipes
Western Blotting
10x Transfer Buffer
- Tris, ultra pure: 15.2 g
- Glycine, ultra pure: 72.1 g
- SDS, ultra pure: 5.0 g
- Ultra pure water to 500 ml
Store at 4°C.
1x Transfer Buffer
- 10x Transfer Buffer: 50 ml
- Methanol: 100 ml
- Distilled water: 350 ml
Make fresh for each use.
TBST
- Tris, ultra pure: 6.1 g
- NaCl: 8.68 g
- Tween 20: 500 µl
Adjust the volume to 1 liter with distilled water. Adjust pH to 8.0 with HCL. Store at 4 - 25°C.
Stripping Buffer
- Glycine, ultra pure: 15 g
- SDS, ultra pure: 1 g
- Tween 20: 10 mL
Adjust the volume to 1 liter with distilled water. Adjust pH to 2.2.
SDS PAGE
20x Running Buffer
- Tricine, ultra pure: 71.7 g
- Tris, ultra pure: 72.6 g
- SDS, ultra pure: 10.0 g
- Sodium bisulfite: 2.5 g
- Ultra pure water to 500 ml
Store at 4°C. For 1x Running Buffer, add 10 ml of 20x Running Buffer to 190 ml of distilled water.
6x Laemmli (Sample) Buffer
- 1 M Tris-Cl, ultra pure: 3.75 mL
- Glycerol: 6 mL
- SDS, ultra pure: 1.2 g
- DTT: 0.93 g
- Bromophenol blue: 6 mg
Adjust the volume to 10 mL with distilled water. Store at -20 °C in 0.5 mL aliquots.
Coomassie Brilliant Blue Solution
- Coomassie Brilliant Blue: 1.25 g
- Methanol: 500 mL
- Acetic acid: 100 mL
- Distilled water: 400 mL
20x MOPS SDS Running Buffer
- MOPS, ultra pure: 209.3 g
- Tris, ultra pure: 121.1 g
- SDS, ultra pure: 20 g
- EDTA: 6 g
Adjust the volume to 1 liter with distilled water
20x MES SDS Running Buffer
- MES, ultra pure: 195.2 g
- Tris, ultra pure: 121.1 g
- SDS, ultra pure: 20 g
- EDTA: 6 g
Adjust the volume to 1 liter with distilled water.
General Use
10x PBS
- NaCl: 8 g
- KCl: 0.2 g
- Disodium phosphate: 1.44 g
- Monopotassium phosphate: 245 mg
Adjust the volume to 1 liter with distilled water. Adjust pH to 7.4.
Agarose Gel Electrophoresis
10x DNA loading buffer
- Glycerol: 3.9 mL
- SDS, ultra pure: 50 mg
- EDTA 0.5 M: 200 µL
- Bromophenol blue: 0.025 g
- Xylene cyanol: 0.025 g
Adjust the volume to 10 mL with distilled water.
50x TAE
- Tris, ultra pure: 242 g
- Acetic acid: 57.1 mL
- EDTA 0.5 M: 100 mL
Adjust the volume to 1 liter with distilled water.
10x TBE
- Tris, ultra pure: 108 g
- Boric acid: 55 g
- EDTA 0.5 M: 40 mL
Adjust the volume to 1 liter with distilled water.
Southern and Northern Blot
20x SSC Buffer
- NaCl: 175.3 g
- Sodium citrate: 88.2 g
Adjust the volume to 1 liter with distilled water. Adjust pH to 7.0 with HCl.
20x SSPE Buffer
- Sodium citrate: 175.3 g
- Monosodium phosphate: 27.6 g
- EDTA: 7.4 g
Adjust the volume to 1 liter with distilled water. Adjust pH to 7.4 with NaOH.
Sedimentation
Sucrose Gradient Buffer
- HEPES, ultra pure: 0.691 g
- Potassium acetate: 1.47 g
- Magnesium acetate: 0.214 g
- β-Mercaptoethanol: 800 µL
Adjust the volume to 100 mL with distilled water.
Sucrose Stock Solution
- HEPES, ultra pure: 0.691 g
- Potassium acetate: 1.47 g
- Sucrose: 60 g
Adjust the volume to 100 mL with distilled water.
Immunoprecipitation
RIPA buffer
- NaCl: 818 mg
- Tris, ultra pure: 121 mg
- EDTA: 29 mg
- EGTA, ultra pure: 19 mg
- Triton X-100: 1 g
- SDS, ultra pure: 0.1 g
- Sodium deoxycholate: 0.1 g
Adjust the volume to 100 mL with distilled water. Add 1 mM PMSF before each use