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Team: University of Chicago/Notebook/SDS-PAGE
From 2008.igem.org
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Materials | Materials | ||
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polyacrylamide gel | polyacrylamide gel | ||
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1X SDS buffer (6.05g Tris, 28.83g glycine, 2g SDS per liter) | 1X SDS buffer (6.05g Tris, 28.83g glycine, 2g SDS per liter) | ||
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Benchmark Protein Ladder | Benchmark Protein Ladder | ||
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Denatured protein samples | Denatured protein samples | ||
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Commassie blue (50% methanol, 10% acetic acid, 0.2% commassie) | Commassie blue (50% methanol, 10% acetic acid, 0.2% commassie) | ||
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Destain solution (10% methanol, 10% acetic acid) | Destain solution (10% methanol, 10% acetic acid) | ||
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1. Take the comb out of the prepared polyacrylamide gel. Place the gel in a holder and attach the holder to the PAGE apparatus. | 1. Take the comb out of the prepared polyacrylamide gel. Place the gel in a holder and attach the holder to the PAGE apparatus. | ||
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2. Insert the apparatus in the tank and pour 1X SDS buffer in the tank and inside the apparatus. | 2. Insert the apparatus in the tank and pour 1X SDS buffer in the tank and inside the apparatus. | ||
3. Pipette 10ul of Benchmark ladder in one lane of the gel | 3. Pipette 10ul of Benchmark ladder in one lane of the gel | ||
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4. Pipette desired amounts of protein samples in other lanes of the gel. | 4. Pipette desired amounts of protein samples in other lanes of the gel. | ||
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5. Put the lid on and connect the cables to the power supply | 5. Put the lid on and connect the cables to the power supply | ||
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6. Set the power supply to 80V. Run for 20 minutes or until the dye front passes the stack. | 6. Set the power supply to 80V. Run for 20 minutes or until the dye front passes the stack. | ||
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7. Increase the voltage to 120V and continue running until the dye front reaches the end of the gel. | 7. Increase the voltage to 120V and continue running until the dye front reaches the end of the gel. | ||
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8. Cut off the power supply and remove gel from the apparatus. | 8. Cut off the power supply and remove gel from the apparatus. | ||
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9. Stain gel in commasie blue stain for 1h | 9. Stain gel in commasie blue stain for 1h | ||
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10. Destain in destain solution for 30min. | 10. Destain in destain solution for 30min. | ||
| - | <<Protocol acquired from | + | <<Protocol acquired from https://2007.igem.org/Melbourne/SDS_PAGE>> |
Latest revision as of 21:09, 12 September 2008
SDS-PAGE Protocol
Materials
polyacrylamide gel
1X SDS buffer (6.05g Tris, 28.83g glycine, 2g SDS per liter)
Benchmark Protein Ladder
Denatured protein samples
Commassie blue (50% methanol, 10% acetic acid, 0.2% commassie)
Destain solution (10% methanol, 10% acetic acid)
Procedures
1. Take the comb out of the prepared polyacrylamide gel. Place the gel in a holder and attach the holder to the PAGE apparatus.
2. Insert the apparatus in the tank and pour 1X SDS buffer in the tank and inside the apparatus. 3. Pipette 10ul of Benchmark ladder in one lane of the gel
4. Pipette desired amounts of protein samples in other lanes of the gel.
5. Put the lid on and connect the cables to the power supply
6. Set the power supply to 80V. Run for 20 minutes or until the dye front passes the stack.
7. Increase the voltage to 120V and continue running until the dye front reaches the end of the gel.
8. Cut off the power supply and remove gel from the apparatus.
9. Stain gel in commasie blue stain for 1h
10. Destain in destain solution for 30min.
<<Protocol acquired from https://2007.igem.org/Melbourne/SDS_PAGE>>
