LumiBlue™ ECL Extra Solution for Western Blotting
LumiBlue™ ECL Extra is a solution with a high level of signal intensity and is effective for qualitative and quantitative detection across a wide range of protein concentrations. The solution is sensitive enough to detect 500fg of protein per band and has a signal duration of up to 12 hours. The working solution is stable for up to three days after activation, allowing convenient experiments spanning multiple days and minimising waste.
LumiBlue™ ECL Extra is at least 20 times more intense than standard ECL based on phenolic enhancers, allowing lower antibody concentrations and more blots per batch of antibody while still retaining the same sensitivity. This kit provides a comparable level of sensitivity to Pierce™ ECL Plus (Thermo Fisher Scientific) and discontinued Amersham™ ECL Plus (GE Healthcare) with the same antibody dilutions, and does not require protocol reoptimization or changes from these kits.
Benefits of LumiBlue™ ECL Extra Include:
- Linear response – Novel acylation catalyst provide stable signal over a wide range of protein concentrations
- High sensitivity – Detect protein levels as low as 500fg per band with next generation enhancers
- Long lasting signal –Perform multiple exposures with less signal loss
- Low background – Optimized formulation for lower backgrounds
- Bright bands – Equal or better brightness compared to other ECL solutions
- Standard protocols – Direct replacement of other ECL solutions with no protocol reoptimization or changes
- Long shelf life – One year shelf life at 4°C, convenient for infrequent Western blotters.
ECL kits are based on the enzymatic oxidation of luminol with peroxide, which produces light, usually through horseradish peroxidase (HRP). This method is employed by most traditional ECL kits. However, LumiBlue™ improves upon standard ECL kits by using a patented next generation electron mediator coupled with a novel acylation catalyst to improve performance (called enhancers).
The combination of these unique modifications to the ECL process increases reaction efficiency resulting in increased light intensity and less signal decay. Different LumiBlue™ kits are extensively optimized for specific antibody dilutions and protein levels giving higher intensity, longer signal stability and wider linearity over traditional ECL kits. This allows for more flexible and cost effective experimental design.
LumiBlue™ ECL solutions are enhanced by a novel acylation catalyst, resulting in a more linear response to HRP levels than conventional ECL solutions. This increases the range of antibody concentrations at which reliable measurement can occur. When appropriate antibodies are used protein levels as low as 50 fg (5×10-14g) per band can be detected with LumiBlue™ ECL Extreme.
Three Day Working Solution
LumiBlue™ ECL solutions are activated by mixing the provided solutions in a 1:1 ratio. This allows smaller volumes to be prepared with LumiBlue™ than with other solutions that require asymmetric ratios such as 40:1. Convenient volumes can be prepared quickly and easily. Once mixed, the ECL Extra working solution is stable for up to three days, permitting larger batches to be made and used for a number of experiments across multiple days, saving time and ensuring that exactly the same solution is used for each blot.
*Antibodies diluted from 1mg / ml stock solution.
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