Buffer Recipes

1x Transfer Buffer

• 10x Transfer Buffer: 50 ml
• Methanol: 100 ml
• Distilled water: 350 ml

Make fresh for each use.

Blocking Buffer

• Non-fat milk powder: 50 g
• TBST: 1 l

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: 1g
• Tween 20: 10 mL

Adjust the volume to 1 liter with distilled water. Adjust pH to 2.2.

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.

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.

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.

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.

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.

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