Blue Native Polyacrylamide Gel Electrophoresis: A Non-Denaturing Separation Technique for Analysis of Intact Mitochondrial Respiratory Chain Complexes

To perform the blue native gel electrophoresis, first, add the blue cathode buffer to the gel cassette. Use a pipette to wash and fill the wells with the blue cathode buffer. Then, load 5 to 30 micrograms of protein samples into the wells. Gently fill the gel cassette to the top with the blue cathode buffer, and the tank with the anode buffer.

First, run the gel at a constant voltage of 40 volts for 15 minutes. Then, increase the voltage to 80 volts. Run the gel until the dye reaches 2/3rds of the gel length. Replace the blue cathode buffer with the cathode buffer, and continue electrophoresis until the dye-front has run off the gel.