Protocol for: Fairbanks staining technique for improved protein visualization on native/SDS-PAGE gels
Abstract or Description:
THIS IS AN AWESOME COOMASSIE STAINING METHOD!
Adapted from : Fairbanks G. et al (1971) Electrophoretic analysis of the major polypeptides of human erythrocyte membrane. Biochemistry, 10: 2606-2617
As little as 5ng of protein can be stained and observed. This technique provides better resolution than traditional Coomassie staining. As with Coomassie staining, solutions A, B and C can be re-used.
0.05% Coomassie R-250 + 25% isopropanol + 10 % acetic acid
0.005% Coomassie R-250 + 10% isopropanol + 10% acetic acid
0.002% Coomassie R-250 + 10% acetic acid
10% acetic acid
1. Transfer gel into plastic container and add 100mL of Fairbanks A solution. Microwave on high for 2 minutes, or until solution reaches boiling point.
2. Remove from microwave and allow to “cool” for 5 minutes at room temp, with gently shaking. Discard solution and briefly rinse gel in distilled water and discard water.
3. Add 100mL of Fairbanks B solution and repeat steps 1 and 2.
4. Add 100mL of Fairbanks C solution and repeat steps 1 and 2.
5. Finally add Fairbanks D solution (destaining solution) and repeat steps 1 and 2 until clear background is obtained.
Citation: Fairbanks G. et al (1971) Electrophoretic analysis of the major polypeptides of human erythrocyte membrane. Biochemistry, 10: 2606-2617