
What is Western Blotting?
Your common Western blotting questions, answered. Western blotting is a widely used analytical technique that can identify one or more specific proteins in a complex
Nucleic acid and protein gels are a fundamental part of lab life. Gel documentation is a method used to record and measure labeled nucleic acid and protein in various types of media, like acrylamide, agarose, or cellulose. DNA or RNA is usually stained by ethidium bromide, which intercalates into the nucleic acids and can be visualized with UV light. Safer dyes for staining DNA (SYBR® Safe, SYBR® Gold, SYBR® Green) are also growing in popularity, and can imaged with blue light.
Protein purification procedures often rely on verifying purity by SDS-PAGE and subsequent gel staining (Coomassie blue or silver stain) to check for contaminating protein bands. Gel documentation system are able to record and measure labeled protein and nucleic acid present across different media, such as cellulose or agarose.
Many stains, including TotalStain Q, AzureRed, Coomassie Fluor™, Deep Purple™, SYPRO® Ruby, SYPRO® Red, SYPRO® Tangerine, and more can be imaged with advanced digital imagers, such as the new chemiSOLO, or Azure Imaging Systems.
Azure Imagers are the perfect solution for high-performance gel documentation and analysis of gels. The Azure 200 has the imaging capabilities for all of these applications. With a dual-wavelength 302 nm and 365 nm UV transilluminator, ethidium bromide-stained DNA gels can be imaged in a fraction of a second. For excising bands from a gel, the UV transilluminator platform can be pulled out.
The Azure 400 and Azure 600 imagers, as well as the Azure Sapphire FL Biomolecular Imager, open up the ability to document protein gels stained with a wide variety of fluorescent protein stains that fluoresce in the visible spectrum.
Gel documentation systems (also called gel docs, or imagers) serve as enclosed darkrooms to block external light from entering the inside. These instruments are used to image gels, Western blots, tissues, arrays, and more. Gel docs usually contain UV transillumination that excites dyes from beneath the area the sample is placed. Light travels through an emission filter, to a high-resolution camera where an image of the sample is collected.
Azure Imaging Systems use CCD cameras with fast lens options, auto detection, and 4.8 OD dynamic range to capture images. Learn more
Polyacrylamide gels form a crosslinked, mesh-like matrix through which proteins migrate with the application of current. The matrix functions like a sieve, allowing smaller, low molecular weight proteins to migrate quickly through the gel, while large, high molecular weight proteins tend to progress more slowly. When the electrophoresis is finished, proteins in each lane will be separated into a continuum, with the highest molecular weight proteins remaining near the loading site and the lowest molecular weight proteins reaching the opposite end of the gel. Read 6 tips on troubleshooting band separation
Proteins travel effectively through the gel matrix based exclusively on size.
FREE WESTERN BLOT eBOOK
A portable, personal chemiluminescent Western blot imager controlled by mobile device
Flexible gel docs for Western blots, DNA gels, and more
Laser and CCD-based scanning system for in-cell Westerns, Western blots, gels, and more
Looking for a full list of applications?
Your common Western blotting questions, answered. Western blotting is a widely used analytical technique that can identify one or more specific proteins in a complex
We’ve all been there… Non-specific bands are a perennially frustrating problem in Western blotting. Here are some of the different reasons you might be getting
Chemiluminescent detection depends on an enzymatic reaction so timing and the amount of both enzyme and substrate used have important effects on data quality. Light
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