SDS sample buffer

Purpose

• For preparation and loading of protein samples onto a gel for SDS-PAGE analysis.

Preparation

To make 10 mL of 4X stock:

• 2.0 ml 1M Tris-HCl pH 6.8
• 0.8 g SDS
• 4.0 ml 100% glycerol
• 0.4 ml 14.7 M β-mercaptoethanol
• 1.0 ml 0.5 M EDTA
• 8 mg bromophenol Blue

Final concentrations (1X):

• 50 mM Tris-HCl pH 6.8
• 2% SDS
• 10% glycerol
• 1% β-mercaptoethanol
• 12.5 mM EDTA
• 0.02 % bromophenol blue

Use of loading buffer

Dilute protein sample 1:3 into 4X sample loading buffer. Load on acrylimide gel in SDS-PAGE buffer.

Safety

β-mercaptoethanol is a severe irritant and is readily absorbed through the skin. SDS is a respiratory irritant in solid form and a mask should be worn while weighing it.

• β-mercaptoethanol MSDS

References

• Szostak Lab SDS-PAGE