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Team:Calgary/Sandbox/Notebook/Protocols/GelShiftAssay
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Revision as of 06:15, 18 September 2013
Gel Shift Assay
Reagents and Materials
10x Buffer- 120mM Tris Cl
- 600mM KCl
- 20mM DTT
- 0.5% NP-40
- 1mg/mL BSA
- 50% Glycerol
- 50 mM MgCl2
- 2mM EDTA
- Fill up to desired volume with water
- pH = 7.5
- 1.3% agarose gel
- DNA of interest
- Purified protein sample
Protocol
- Perform a Bradford Assay to the protein concentration
- Add 2µL of 10X Buffer, 55pM of DNA, 0.01 to 2500nM of Protein, and fill up to 20µL with ddH2O. Mix thoroughly
- Incubate for 1 hour at room temperature in the dark
- Place at 4°C for 30 minutes
- Run on a 1.3% agarose gel at 50V
