Chemiluminescent detection uses an enzyme to catalyze a reaction that results in the production of visible light. Detection is achieved by either exposing the blot to X-ray film or acquiring the image directly in a chemiluminescence- compatible digital imaging system, usually equipped with highly-cooled CCD cameras to avoid electronic noise. Reprobing is possible with chemiluminescent substrates.
There are a variety of chemiluminescent substrates offering researchers different levels of sensitivity of detection. Traditional or low-sensitivity substrates allow protein detection at the picogram level. While these substrates may be appropriate for routine applications, they cannot detect low abundance proteins. New, high-sensitivity substrates, such as Immobilon® substrates
, allow visualization of proteins at the femtogram level. However, use of these powerful substrates often requires optimization of primary and secondary antibody concentrations.
Luminata™ Western HRP Substrates
Chemiluminescent HRP (horseradish peroxidase) substrates (also known as ECL reagents) are the most sensitive reagents used in the detection of Western blots.
The Luminata™ Western HRP Substrates are a family of three premixed HRP substrates, which offer several advantages over other detection reagents. Pour directly onto the Western membrane without worrying about pipetting error. The Luminata™ Classico, Crescendo and Forte
substrates cover a broad range of sensitivities.
Advantages of Luminata™ Chemiluminescent Detection Substrates
- Premixed for convenience and to avoid cross contamination of the reagents.
- Broad range of sensitivities to satisfy all detection needs.
- Consistent results with less pipetting error.
- Stable at 4 °C or room temperature so you can store it on your own bench.
- Best detection performance relative to competition.
- Compatible with both PVDF and nitrocellulose membranes, as well as all commonly used buffers and blocking reagents.
SignalBoost™ Immunoreaction Enhancer Kit
|Detecting proteins in a Western can be difficult for multiple reasons (low protein abundance, low affinity antibody, epitope availability, etc.). SignalBoost™ mmunoreaction Enhancer can amplify your signals so you can get your data more quickly and spend less time troubleshooting.
Features and benefits:
- Increases signal: minimizes background in immunoblotting, dot blotting and ELISA
- Use with nitrocellulose or PVDF membranes
- Compatible with chemiluminescent or colorimetric detection systems
- Ready-to-use format, no dilution necessary
- Cost-effective: use at least 20-fold less primary antibody in western blotting compared with standard protocols 20 miniblots/1600 cm 2
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ReBlot™ Antibody Stripping Solution
|This quick stripping reagent is the product of choice for regenerating Western blots. ReBlot™ Plus reagents efficiently strip probed blots of bound antibodies. ReBlot™ Plus reagents are available in two formulations, "Mild" and "Strong".