Melbourne/SDS PAGE
From 2007.igem.org
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#Increase the voltage to 120V and continue running until the dye front reaches the end of the gel. | #Increase the voltage to 120V and continue running until the dye front reaches the end of the gel. | ||
#Cut off the power supply and remove gel from the apparatus. | #Cut off the power supply and remove gel from the apparatus. | ||
| - | # | + | #Stain gel in commasie blue stain for 1h |
| + | #Destain in destain solution for 30min. | ||
=====Equipement Required===== | =====Equipement Required===== | ||
Revision as of 12:24, 29 September 2007
<Return to list of protocols> <Team home page>
- Applications:
- Separation of proteins on a gel
- Western blots
- Time to complete protocol:
- Lab time: 15min.
- Waiting time:2h
- Approximate cost of materials: $
Contents |
Method from primary and secondary reagents
Primary & secondary Reagents Required including controls
- polyacrylamide gel
- 1X SDS buffer
- Benchmark Protein Ladder
- Denatured protein samples
Method including controls
- Take the comb out of the prepared polyacrylamide gel. Place the gel in a holder and attach the holder to the PAGE apparatus.
- Insert the apparatus in the tank and pour 1X SDS buffer in the tank and inside the apparatus.
- Pipette 10ul of Benchmark ladder in one lane of the gel
- Pipette desired amounts of protein samples in other lanes of the gel.
- Put the lid on and connect the cables to the power supply
- Set the power supply to 80V. Run for 20 minutes or until the dye front passes the stack.
- Increase the voltage to 120V and continue running until the dye front reaches the end of the gel.
- Cut off the power supply and remove gel from the apparatus.
- Stain gel in commasie blue stain for 1h
- Destain in destain solution for 30min.
Equipement Required
- Pipette and tips
- SDS PAGE apparatus
- Power source
