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Fairbanks staining technique for improved protein visualization on native/SDS-PAGE gels

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Abstract or Description:

Fairbanks Staining

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.

Solutions:

Fairbanks A:

0.05% Coomassie R-250 + 25% isopropanol + 10 % acetic acid

Fairbanks B:

0.005% Coomassie R-250 + 10% isopropanol + 10% acetic acid

Fairbanks C:

0.002% Coomassie R-250 + 10% acetic acid

Fairbanks D:

10% acetic acid

 

Protocol:

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
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Posted By: Robyn_L_K on 5/28/2013 2:07:41 PM
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