The script that has been used to create the video shown above can be downloaded as excel script file.
Soak 1 stack of transfer stacks in 35 ml Blot buffer.
Place the transfer stack in the Turbo-Blot tray.
Keep the membrane between the blue papers as much as possible to avoid protein contamination. Do not touch the membranes with bare hands: wear gloves and use tweezers.
Do not touch the membranes with bare hands: wear gloves and use tweezers.
Carefully remove the top glass plate from the gel.
Remove the stacking gel (the part of the gel where the slots are) and gently place the gel on the membrane.
Repeat this step for the 2nd gel.
Start the 7-minute transfer program.
Remove the tray from the machine and check if you can see the protein ladder on the membrane (NOTE: On which side are your proteins ?!)
What is the correct way to assemble a transfer stack? Start with the tray block on top, and add the others in the right order downwards.
Now you know how to build up your transfer stack and you know how the charge has influence on the process, can you explain where the positive and negative pool are located?
Proteins are [a].
This means the tray contains the [b] pool, and the lid contains the [c] pool.
Wat gebeurt er bij activatie van het PVDF membraan?
Which is the correct way to assemble a transfer stack? Fill in the correct answer:
Negative Terminal (anode) – [a] – [b] – [c] – [d] – Positive Terminal (cathode)
Think about which item you use more than once. Which way does the current flow? And on which side of the membrane, gel or transfer stack is the protein?
Correct answer = Negative Terminal (anode) – [a=transfer stack] – [b=gel] – [c=PVDF membrane] – [d=transfer stack] – Positive Terminal (cathode)