To make 30 mL of 2X SDS sample buffer (with 2-β-ME):

Ingredients:

• 1 M Tris-HCl (pH 6.8): 7.5 mL
• 10% SDS: 12 mL
• Glycerol (100%): 6 mL
• Bromophenol blue (0.1%): 300 µL
• 2-β-Mercaptoethanol (100%): 1.5 mL
• Distilled water: To 30 mL

Steps:

Ready to use for SDS-PAGE!

Here is the recipe we use in our lab; feel free to adapt it to suit your specific needs.

2X SDS-PAGE Sample Buffer Recipe (30 mL Total Volume)

Ingredients:

0.125 M Tris-HCl → 1.88 mL of 1M Tris-HCl

4% (w/v) SDS → 1.2 g SDS

20% (v/v) Glycerol → 6 mL Glycerol

0.01% (w/v) Bromophenol Blue → 3 mg Bromophenol Blue

Adjust volume to 30 mL with distilled, deionized water. The pH of this solution is 6.8.

10% (v/v) β-Mercaptoethanol (β-Me) (add fresh before use) → 3 mL β-Me

Preparation Instructions:

Dissolve Tris-HCl, SDS, and Bromophenol Blue in approximately 25 mL of distilled, deionized water.

Add Glycerol and mix thoroughly.

Adjust the final volume to 30 mL with distilled, deionized water.

Add 10% β-Mercaptoethanol just before use if reducing conditions are needed.

Aliquot and store at -20°C. If β-Me is added, store at 4°C for short-term use.

🔹 Usage: Mix 1:1 with your protein sample before boiling at 95°C for 5 minutes.