Team:Macquarie Australia/WetLab/Protocols/SDSPAGE

SDS-PAGE

• Re-suspend pelleted bacterial cells in 200µL of Milli-Q-water
• Transfer 50 µL of suspension into new Eppendorf tubes and combine with 50ul of 2xTruSep sample buffer.
• Shear the cells using a Hamilton syringe.
• Centrifuge the preparation for 3minutes @ 13,000 rpm.
• Load 20 µL of the supernatant into gel.
• Conduct electrophoresis at a constant voltage (200V) for 1 hour.
• Coommassie Stain for ~30 minutes.

Figure 1. Protein bands from SDS-PAGE, visualized under UV light.