Transfering a Protein Gel

Transfering a Protein Gel

Transferring a Protein Gel

1. Once run is complete (blue is running out of gel) switch off power supply
2. Place transfer buffer into a container (found on drying rack)

Transfer buffer: 28.8g glycine, 6.07g tris base, 200 ml methanol

1. Build your sandwich in transfer buffer (two sandwiches)

• White side of sandwich holder down
• One black mesh pad
• 2 layers of filter paper
• One piece of nitrocellulose membrane (not PVDF) fitted to gel size
• Your gel

To remove gel: take glass plates carefully out of the glass plate holder Remove the smaller glass plate by slowly turning one spacer towards you, lifting the plate and making the gel stick to the larger plate Take the larger plate into the solution with the sandwich and use a razor blade to carefully pry the gel from the larger glass plate Once it is onto the membrane in solution, adjust so gel is centered on membrane

• 2 layers of filter paper
• One black mesh pad

1. Black side of sandwich holder down, and close tight
2. Place both sandwich holders into transfer apparatus with both black sides facing the black wall of the transfer apparatus. Put the entire apparatus into the transfer chamber
3. Place an ice block (from the freezer) into the chamber and fill with transfer buffer to the top
4. Place the chamber top on, and switch on the power supply
5. Transfer the gel at 500 volts/ 200 milliamps on the bench, overnight