Jacobs:Protocol Protein Gel Staining Using Coomassie Blue Protein Stain

Materials

• Polyacrylamide pre-cast gel
• SimplyBlue™ Coomassie protein stain, 1x pre-mixed solution (Coomassie blue dye binds to proteins (arginine, the aromatic amino acids, and histidine) allowing protein bands to be visualized in polyacrylamide gels.)
• Plastic container
• Gel separator

Procedure

1. After electrophoresis, remove the gel from the gel box and plates using a gel separator
2. Place it in a clean plastic container and cover with ultrapure water
3. Microwave the gel on high for 1 minute until the solution almost boils (do not overheat)
4. Shake the gel on a shaker for 2 minute and discard the water
5. Repeat steps 3 and 4 two more times
6. After the last wash, add 30 ml of SimplyBlue™ Coomassie protein stain and microwave on high for 45 seconds to 1 minute until the solution almost boils
7. Shake the gel on a shaker for 10 minutes
8. Wash the gel in 100 ml of ultrapure water for 10 minutes on a shaker
9. Add 20 ml of 20% NaCl for at least 5 minutes
10. Examine gel on a white paper towel for blue-stained protein bands
11. Discard the gel in the Biohazard waste bin

Contact

• History: CRJ-ABC, Last updated: 8/3/07

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