Gel Shift Assay

Reagents and Materials

• 1.3% agarose gel
• DNA of interest
• Purified protein sample

10x Buffer

• 120mM Tris Cl
• 600mM KCl
• 20mM DTT
• 0.5% NP-40
• 1mg/mL BSA
• 50% Glycerol
• 50 mM MgCl2
• 2mM EDTA
• ddH₂O (deionized water)
• pH = 7.5

Protocol

1. Perform a Bradford Assay to the protein concentration
2. Add 2µL of 10X Buffer, 55pM of DNA, 0.01 to 2500nM of Protein, and fill up to 20µL with ddH₂O. Mix thoroughly
3. Incubate for 1 hour at room temperature in the dark
4. Place at 4°C for 30 minutes
5. Run on a 1.3% agarose gel at 50V