Learning Objectives:

• 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.

SOLUTION

Protein gel electrophoresis can be followed by the addition of antibodies to identify specific proteins. This technique, described in the steps below, is called the Western blot.

STEP BY STEP

Step 1. Polyacrylamide gel electrophoresis (PAGE), is used to separate proteins on an electrophoresis gel.
Step 2. Protein antigens in the gel are transferred to a nitrocellulose membrane, immobilizing them.
Step 3. The nitrocellulose membrane is exposed to a primary antibody produced to bind to the protein of interest.
Step 4. A second antibody equipped with a molecular beacon (i.e., something that makes it visible) is added to bind to the first antibody (primary antibodies can also be tagged with a molecular beacon in some cases).….Please click the icon below to access entire solution at $10