Membranes for Western Blotting

Western blotting is an analytical technique in molecular biology and is the basis of protein analysis. It is often used to study and characterize protein post-translational modification, protein identification, and protein production verification. Simple and effective Western blotting has applications in many environments, including basic scientific research, biopharmaceutical production, forensic medicine, and diagnosis. The successful outcome may depend on several factors, including the best membrane selection for the required detection method and protocol.

High-quality membranes are essential to assess the relative abundance of proteins for analysis. Alfa Chemistry can help you obtain reliable and reproducible Western blot results. Our transfer film series supports multiple detection methods without compromising quality.

Different Western Blot Detection Methods

Although the steps of Western blotting are basically the same, the detection methods are different.

Figure.1 Steps involved in western blotting procedure

The most commonly used detection methods include radioactive labeling, fluorophores, chromogenic and chemiluminescent enzymatic reactions. Each method has its own advantages and precautions:

• Radiolabeled probes support the direct detection of proteins through X-ray film; due to radioactivity, safety requirements need to be strengthened.
• The color reaction does not require special detection equipment, and the color formed on the film can be observed with the naked eye.
• Chemiluminescence-reaction uses photosensitive equipment or materials to process and further analyze the Western blot results.
• Fluorescent probes used as detection methods allow multiple and simultaneous detections of multiple proteins with different molecular weights.

Both qualitative and quantitative results depend on the difference between the background and the target protein signal.

Transfer Film Optimized for Low Noise and Clear Results

Alfa Chemistry offers a variety of transfer membranes optimized for Western blotting applications. The sensitive nature of our membrane produces high-resolution results, low background, and burn-through.

Our PVDF transfer membrane A has been optimized for fluorescence detection to ensure that it does not interfere with the very low background of protein detection and analysis when exposed to fluorescence. When using fluorescence detection methods, autofluorescence from standard Western transfer membranes can mask specific signals, especially at lower wavelengths.

Our nitrocellulose transfer membrane is a pure nitrocellulose carrier-free medium, compatible with a variety of detection systems, and has a high binding capacity to nucleic acids and proteins.

For more information, please refer to the following selection guide.