Coomassie Staining and Destaining
1. Remove SDS-PAGE gel from glass and rinse once in ddH2O in a suitable container with a lid. Try not to use a container much larger or much smaller then the gel.
2. Add enough Coomassie Stain to cover the gel by 1/2 inch (~ 1.5 cm).
3. Microwave on high power for 40 seconds to 1 minute (until the Coomassie Stain boils).
4. Incubate the gel in the Coomassie stain for 5 to 10 minutes on a rocking table. If you did not microwave the Coomassie/gel, incubate for at least 1 hour.
5. Pour off the Coomassie Stain. The Coomassie Stain can be recycled a couple of times by filtering it.
6. Rinse twice in ddH2O or used Destain solution to remove Coomassie Stain from the container.
7. Add fresh Destain solution to cover the gel by 3/4 inch (~ 2 cm).
8. Tie Kimwipes in a simple knot and place 4 of them in the Destain solution around the gel. Try to avoid laying the Kimwipes on the gel as this will cause an uneven destaining.
9. Microwave on high power for 40 seconds to 1 minute (until the Destain boils).
10. Incubate the gel in the Destain solution for 10 minutes on a rocking table. If you did not microwave the Coomassie/gel, incubate for at least 1 hour.
11. Discard the stained Kimwipes and replace with fresh knotted Kimwipes.
12. Incubate a second time for 10 minutes to overnight on a rocking table. Stop whenever the level of destaining is sufficient for you. Microwave again to speed up the process.
13. The used Destain solution can be recycled a couple of times by storing it in a sealed container with sponges or Kimwipes to remove all traces of Coomassie Stain.
Coomassie Stain 1 L:
0.1% Coomassie R250, 10% acetic acid, 40% methanol
1. Add 100 ml of glacial acetic acid to 500 ml of ddH2O.
2. Add 400 ml of methanol and mix.
3. Add 1g of Coomassie R250 dye and mix.
4. Filter to remove particulates (a coffee filter works great for this and is cheap)
5. Store at room temperature in a sealable container.
De-Stain for Coomassie 1 L:
1. Add 100 ml of glacial acetic acid to 700 ml of ddH2O.
2. Add 200 ml of methanol and mix.
3. Store at room
2. Add enough Coomassie Stain to cover the gel by 1/2 inch (~ 1.5 cm).
3. Microwave on high power for 40 seconds to 1 minute (until the Coomassie Stain boils).
4. Incubate the gel in the Coomassie stain for 5 to 10 minutes on a rocking table. If you did not microwave the Coomassie/gel, incubate for at least 1 hour.
5. Pour off the Coomassie Stain. The Coomassie Stain can be recycled a couple of times by filtering it.
6. Rinse twice in ddH2O or used Destain solution to remove Coomassie Stain from the container.
7. Add fresh Destain solution to cover the gel by 3/4 inch (~ 2 cm).
8. Tie Kimwipes in a simple knot and place 4 of them in the Destain solution around the gel. Try to avoid laying the Kimwipes on the gel as this will cause an uneven destaining.
9. Microwave on high power for 40 seconds to 1 minute (until the Destain boils).
10. Incubate the gel in the Destain solution for 10 minutes on a rocking table. If you did not microwave the Coomassie/gel, incubate for at least 1 hour.
11. Discard the stained Kimwipes and replace with fresh knotted Kimwipes.
12. Incubate a second time for 10 minutes to overnight on a rocking table. Stop whenever the level of destaining is sufficient for you. Microwave again to speed up the process.
13. The used Destain solution can be recycled a couple of times by storing it in a sealed container with sponges or Kimwipes to remove all traces of Coomassie Stain.
Coomassie Stain 1 L:
0.1% Coomassie R250, 10% acetic acid, 40% methanol
1. Add 100 ml of glacial acetic acid to 500 ml of ddH2O.
2. Add 400 ml of methanol and mix.
3. Add 1g of Coomassie R250 dye and mix.
4. Filter to remove particulates (a coffee filter works great for this and is cheap)
5. Store at room temperature in a sealable container.
De-Stain for Coomassie 1 L:
1. Add 100 ml of glacial acetic acid to 700 ml of ddH2O.
2. Add 200 ml of methanol and mix.
3. Store at room