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In this visual protocol series, learn how learn how to perform all phases of a western blot using the most common methods for this assay.
Phase One - Sample Preparation
Learn how to prepare a sample lysate from cell culture.
Phase Two - Protein Electophoresis (SDS-PAGE)
Learn how to load a gel and separate the individual proteins in the sample lysate through electrophoresis.
Phase Three - Membrane Transfer
Learn how to transfer separated proteins from the SDS-PAGE gel into a solid membrane, or blot.
Phase Four - Immunoblotting
Learn how to block the blot, apply a primary antibody specific for our protein of interest, and then a secondary antibody which will recognize the primary antibody.
Phase Five - Detection
Learn how to visualize your protein of interest that was probed with specific antibodies in the previous step. We will demonstrate signal development using the most common, sensitive, and inexpensive detection method, the electrochemiluminescent, or ECL, reaction.
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