Running SDS-PAGE

SDS PAGE

See also SDS-PAGE Protocol

Running SDS-PAGE

1. Make ready the samples to run
2. Make ready the sample running buffer
3. Make ready the gel
4. Have the SDS-PAGE apparatus
5. Place the the gel in the SDS-PAGE stand
6. Place the stand with the gel in the SDS-PAGE apparatus bath
7. fill the space between the gels with running buffer
8. Take off the combs of the gel (take it out gently)
9. Load the sample to each well
10. Do not use the wells at the left and right end of the gel
11. fill the SDS-PAGE apparatus bath with running buffer to the bottom of the gel
12. Place the cover of the SDS-PAGE
    1. N.B Make sure the correct plug is connected to the correct electrode (red goes to red and black goes to black)
13. Plug the cables to the SDS-PAGE apparatus power source
14. Turn of the SDS-PAGE power source
15. Set to constant voltage
16. Use 200 V
17. Press the Run button to start the electrophoresis
    1. The voltage increases to 200 V from 0 V
    2. For 4% staking gel and 12.5 % separating gel the current reaches 50-60 mA and the power 10 -12 W
    3. The current the the power decreases in time but not reach zero (if it reaches zero see the troubleshoot)
    4. For the state type of gel it takes 35 - 40 min

Troubleshoot

1. The power drops to zero

• really old buffer
• the space in between the gels is not filled with buffer to the top