Western Blot Detection Reagents
With a variety of detection techniques for western blotting-chemiluminescence, fluorescence, or colorimetric, to choose from, you can select a technology to match your experimental requirements by clicking “Learn more” below.
Chemiluminescent substrates are popular because they offer several advantages over other detection methods. These advantages have allowed chemiluminescence to become the detection method of choice in most protein laboratories. Key highlights: Greatest sensitivity; Detection and quantitation for a large range of protein concentrations.
Fluorescent reagents are growing in popularity for western blotting because they offer increased time savings over chemiluminescent detection and reduced chemical waste compared to both chemiluminescent or chromogenic detection systems. Key highlights: Multiplexing capability; Direct signal.
Chemiluminescent detection |
Fluorescent detection |
Chromogenic detection |
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| Major advantage | Highest sensitivity | Detect multiple targets at once | Easily visualize blots without equipment |
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| Technology overview | Enhanced chemiluminescent (ECL) HRP and AP substrates providing pictogram to femtogram level detection | Simultaneous detection of multiple proteins on the same blot using a variety of available fluorescent dyes and conjugated 2° antibodies | Direct visualization of your target protein using color detection reagents |
| Signal Source | Indirect signal from enzymatic reaction | Direct signal from fluorophore | Indirect signal from enzymatic reaction |
| Signal Duration | Limited (hours) | Extended (weeks to months) | Extended (weeks to months) |
| Sensitivity | 
Excellent, with a wide variety of substrates available |
Good, may require higher concentration of secondary antibody |
Limited, best for high abundant proteins |
| Consistency | Possible variation between blots, can be mitigated by using high duration substrates | High reproducibility between blots | Possible variation between blots |
| Detection | X-ray film and imaging instruments | Imaging instruments with appropriate filters or lasers | Visual, no instrumentation required |
| Quantitation | Single-channel detection makes normalization challenging | Multiplexing with an internal control makes normalization simpler | Single-channel detection makes normalization challenging |
| Other considerations |
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