MAAD Scientist Technologies

Learn · Western blot

The Overnight Transfer

Three proteins on one membrane, 10, 45 and 250 kDa. The default recipe loses both ends. Then the contrast slider is right there.

Best full screen: the text is small otherwise. Click the game first, then use the arrow keys to choose, Enter to confirm and Esc to leave. It needs a keyboard, and takes a few seconds to load. Open it in its own tab.

What you do

You have run a gel with three proteins in it: a small one, yours, and a big one. Now they have to get out of the gel and onto a membrane, overnight, in the cold room.

You choose the membrane, the buffer, the voltage and which way round the sandwich goes. In the morning you find out what made it across.

Then the blot goes into a figure, and you are handed a contrast slider.

What you find out

  • Why the membrane goes on one particular side of the gel, and what happens when it doesn’t.
  • Why small proteins go straight through one membrane and stay on another.
  • Why the ingredient that helps proteins stick also keeps the big ones from leaving the gel.
  • Why the transfer runs cold, and what heat does to it.
  • What an air bubble looks like the next day.
  • How far you can adjust an image before it stops showing what happened. A faint second band appears in the treated lanes. The slider can make it vanish.

Where the science comes from

The directions come from the manufacturers’ transfer manuals and from the published guidance on what counts as acceptable image adjustment. How much of each protein transfers under each choice is the game’s own model, and the game says so.