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Western Blot Transfer: Prepare for protein detection Virtual Lab

Join Dr. One and two colleagues on their mission to cure cancer. They are using Western blot to compare the level of p53 protein in cancerous and healthy control cells. Can you help Dr. One with the tricky membrane transfer step?

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Western Blot Transfer: Prepare for protein detection

About this Simulation

Air bubbles are the enemy and if you are not careful you may cause a minor explosion! In this simulation you will learn how to set up and troubleshoot the transfer step of Western blot. Dr. One has already separated the proteins by size using gel electrophoresis. Before they can continue to use antibodies to detect the levels of p53 protein, the protein samples must be transferred from the gel to a membrane.

Building a stack

You will use electricity to transfer the proteins from the gel to the membrane. The gel and membrane must stay closely together during the whole procedure. To achieve this, you start by building the transfer stack. Be careful, as a mistake here could ruin your experiment! Luckily Dr. One is on hand to guide you.

Transferring the proteins

Once you have built the stack, you are ready to assemble the tank in which you will use electricity to move the proteins onto the membrane. As your colleagues Jenny and Mario will be able to tell you, even with the perfect stack, there is still the potential to cause a minor fire or other disaster at this point! Thankfully, the process is much quicker in the virtual lab as you do not need to wait for the transfer to complete.

Analyzing your results

Using antibodies to detect proteins is another lengthy process. Before handing your membrane over to Dr. One for this part of the Western blot experiment you want to make sure that you have been successful. You will use ponceau to stain all the proteins on the membrane so you can assess the quality of your transfer. Jenny and Mario have been working alongside you and need some help checking their membranes too. Can you find the best membranes for Dr. One to analyze?

Learning Objectives

At the end of this simulation, you will be able to:

  • Build a transfer stack with the correct orientation

  • Set up a transfer tank to transfer protein from an SDS-PAGE gel to a membrane.

  • Critique the quality of protein transfer using a removable protein-identifying stain.

Techniques in Lab

At the end of this simulation, you will be able to:

  • Western blotting

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