Protein Electrophoresis and Western Blotting

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.

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.

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

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 chemistr
• Sample volume
• Separation type
• Well format
• Separation range
• Number of wells