How to Prepare 10X SDS-PAGE Running Buffer

1. 1Step 1: Add approximately 800 mL of distilled water to a 1L beaker or flask
2. 2Step 2: Add 30.29 g Tris base to the water and stir until completely dissolved
3. 3Step 3: Add 144.13 g glycine gradually while stirring. Continue stirring until fully dissolved (may take 10-15 minutes)
4. 4Step 4: Once Tris and glycine are completely dissolved, slowly sprinkle 10.0 g SDS powder onto the surface while gently stirring to minimize foam formation
5. 5Step 5: Continue gentle stirring until SDS is completely dissolved. Allow any foam to completely subside before proceeding
6. 6Step 6: Transfer solution to 1L volumetric flask and bring to final volume with distilled water
7. 7Step 7: Mix thoroughly by gentle inversion. Avoid vigorous agitation to prevent foam formation

Do not adjust pH. The natural pH should be approximately 8.3 due to the Tris-glycine buffer system. Significant deviation indicates preparation error.

Filter sterilize through 0.22 μm membrane filter. Do not autoclave as high temperature can cause SDS precipitation and degradation.

Wear nitrile gloves, safety glasses, and lab coat. Handle SDS powder in fume hood to avoid inhalation. SDS can cause skin and eye irritation. Avoid creating aerosols during dissolution.

Store at room temperature in tightly sealed container for up to 6 months. SDS may crystallize at low temperatures - warm to room temperature and mix thoroughly before use if crystals form. Protect from contamination.