ECL Extreme solution for Western blotting
LumiBlue ECL Extreme is a solution with extreme signal intensity and is most effective for very dilute, low abundance western blotting applications where maximum sensitivity is essential. The solution is sensitive enough to detect approximately 50 fg of protein per band and has a signal duration of up to 8 hours. ECL Extreme has an intensity 150 times greater than standard ECL substrates and gives bright, clear bands at super-low protein concentrations. This kit provides a comparable level of sensitivity to Supersignal™ West Femto (Thermo Fisher Scientific) Amersham™ ECL Select (GE Healthcare) with the same antibody dilutions and protocol, and does not require protocol re-optimization or changes from these kits.
Benefits of LumiBlue ECL Extreme include:
- Linear response – novel acylation catalyst provide stable signal over a wide range of protein concentrations
- High sensitivity – detect protein levels as low as 50fg per band with next generation enhancers
- Stable solution – working solutions stable for up to 5 days (ECL Express)
- Long lasting signal – perform multiple exposures with less signal loss
- Low background – optimized formulation for lower backgrounds
- Wide detection range – 5 different ECL solutions, suitable for most protein and antibody concentrations
- Bright bands – Equal or better brightness compared to other ECL solutions
- Standard protocols – Direct replacement of other ECL solutions with no protocol re-optimization or changes
- Long shelf life – One year shelf life at 4°C, convenient for infrequent western blotters
LumiBlue 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 optimised 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.
3 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 Pico working solution is stable for up to 3 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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