Polyacrylamide gel electrophoresis (SDS-PAGE), written by Rosa Schier

MES SDS Running Buffer: dissolve one powder bag in 1L of ddH2O (from GenScript)

• To prepare the final sample of a purification (we want 4ug of protein in 20ul of sample = 4ug protein + …H2O + 10ul loading buffer (calculations with concentration))
• Heat up the samples 5min at 95°C
• Put the gel (from Invitrogen) in the tank (be careful to take off the white sticker and take out the comb)
• Put MES SDS Running Buffer and rinse the holes
• Put 10ul of sample in each hole and 5ul of AcuteBand Pre-stained Protein Ladder (from lubio science) (don’t put the ladder in the middle)
• Let it run for 30min at 180V and 400mA + write down the order of the samples
• Take out the gel, put it in QuickBlue Protein Stain (from lubio science) and let it shake slowly
• Rinse the tank with water
• Once the gel is stained rinse with water
• Scan the gel