Protein Electrophoresis and Western Blotting

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What is Western Blot?

Western blot, or western blotting, is a process that detects specific proteins in a complex mixture of proteins extracted from cells or tissues.

Western blot originated in the late 1970s and is now a routine protein analysis technique used extensively by biochemists, molecular biologists, and cell biologists. The method results in both qualitative and semi-qualitative data and is helpful in a wide range of applications, including:

Lyme disease
BSE (Bovine Spongiform Encephalopathy)

The three main steps in the workflow are separation by size, transfer to stable support, and visualization using appropriate primary and secondary antibodies.

Step one: Separation by size

Scientists and technicians use protein gel electrophoresis to separate protein mixtures according to the molecular weight of each component protein.

Step two: Transfer to stable support

After the proteins have been separated, they are transferred to a stable support membrane using a blotting apparatus.

Step three: Visualization with primary and secondary antibodies

In step three, antibodies specific to the protein of interest bind and are detected after the film is developed or digital images are processed.

How to Choose Your Western Blot Equipment

Reliable results may depend on using the proper western blotting protocol specific to your western blot equipment. Blotting kits simplify the purchasing process and include all necessary components to transfer protein, DNA, or RNA samples onto membranes for staining.

Avantor® can deliver individual components from various equipment, products, and supplies to help create a solution that supports your workflow.

What Is Protein Electrophoresis?

Protein electrophoresis is a laboratory technique in which charged protein molecules are separated according to physical factors by an electrical current that forces them to move through a gel. Protein gel electrophoresis is used in many proteomic analyses, including western blot, and helps identify various diseases. Protein electrophoresis is used to diagnose a type of cancer called myeloma, or multiple myeloma. It can also be used to diagnose other conditions, including:

Waldenström macroglobulinemia
Primary amyloidosis
Monoclonal gammopathy of undetermined significance (MGUS)
Diabetes
Anemia
Thyroid problems
Liver diseases
Certain autoimmune diseases
Nutrient absorption inability or poor nutrition

Selecting the Right Protein Gel for Your Research Application

Select the gel that’s best for your protein electrophoresis experiment by matching gel features to your needs. Consider your requirements for the following options:

Sample type
Gel format
Compatible gel tank
Gel chemistry
Sample volume
Separation type
Well format
Separation range
Number of wells

Protein Extraction & Sample Prep

Quality data depends on high-quality sample preparation. Find everything you need to prepare high-integrity protein samples and successfully extract them.