Unique Technology
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.
5 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 Express working solution is stable for up to 5 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
Additional Information
| Signal Intensity |
Standard |
| Protein Quantity |
High Abundance |
| Sensitivity per band |
low pg (~20pg) |
| 1° Ab dilution* |
1:100 - 1:5,000 |
| 2° Ab dilution* |
1:1,000 - 1:15,000 |
| Signal Duration |
16 Hours |
| Working solution stability |
5 days |
| Alternative to |
Pierce™ ECL (Thermo Fisher Scientific) / Amersham™ ECL (GE Healthcare) |
| Storage |
Stable for 1 year at 4 °C. Product is shipped at ambient temperature. |
| Shelf Life |
1 Year from receipt |
Features & Benefits
- • 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 – Optimised 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
Linear Response
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 (5x10-14g) per band can be detected with LumiBlue ECL Extreme.
Brightness and Stability
LumiBlue ECL solutions have higher and steadier light emission produced by second generation chemiluminescent substrates. Each solution is produced with a different enhancer tuned for optimum brightness and stability. This also gives the possibility of using lower levels of primary and secondary antibody. This can considerably reduce the overall cost for Western blot assays. Solutions are activated by mixing two solutions in a 1:1 ratio for simple storage and activation.
Light Emission
LumiBlue ECL solutions use unique patented downstream catalysts and enhancers to greatly increase the brightness and stability of light produced by luminol and HRP. This allows flexible experimental design with increased time between blotting and analysis, as well as the ability to repeat exposures with negligible luminescence loss.
Compatibility with Standard Protocols
LumiBlue ECL kits have been developed to be compatible with standard ECL protocols used by other kits. In most cases experimental design does not need to be changed if switching between another ECL substrate and LumiBlue, but optimisation with LumiBlue ECL solutions may result in the use of more dilute antibodies for similar experiments.
LumiBlue ECL Range for Western Blot Detection
After establishing itself as a leader in protein staining with the InstantBlue single step 15 minute Coomassie stain, Expedeon has now launched its own range of ECL kits for protein detection in Western blotting. The LumiBlue ECL range consists of 5 kits with increasing levels of sensitivity to ensure reliable detection of protein ranging from high abundance expression to extremely dilute samples of protein and antibody. The LumiBlue ECL range has been designed to match existing kits to allow easy transition without protocol re-optimisation or changes to currently used methods. LumiBlue ECL solutions permit the use of less antibody while still obtaining excellent results, which often results in more economical use of precious antibodies.
**Human Transferrin serially diluted (5 to 0.5ng) and electrophoresis performed. The gels were transferred to PVDF membranes, blocked and incubated with different dilutions of rabbit anti-transferrin. After washing, the membranes were incubated with different dilutions of HRP-conjugated goat anti-rabbit antibody. The membranes were washed again and then incubated with substrate. Blots were acquired with ImageQuant LAS 4000 (GEHC) for 300 sec.
Unique Technology
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.
5 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 Express working solution is stable for up to 5 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
Additional Information
| Signal Intensity |
Standard |
| Protein Quantity |
High Abundance |
| Sensitivity per band |
low pg (~20pg) |
| 1° Ab dilution* |
1:100 - 1:5,000 |
| 2° Ab dilution* |
1:1,000 - 1:15,000 |
| Signal Duration |
16 Hours |
| Working solution stability |
5 days |
| Alternative to |
Pierce™ ECL (Thermo Fisher Scientific) / Amersham™ ECL (GE Healthcare) |
| Storage |
Stable for 1 year at 4 °C. Product is shipped at ambient temperature. |
| Shelf Life |
1 Year from receipt |
Features & Benefits
- • 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 – Optimised 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
Linear Response
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 (5x10-14g) per band can be detected with LumiBlue ECL Extreme.
Brightness and Stability
LumiBlue ECL solutions have higher and steadier light emission produced by second generation chemiluminescent substrates. Each solution is produced with a different enhancer tuned for optimum brightness and stability. This also gives the possibility of using lower levels of primary and secondary antibody. This can considerably reduce the overall cost for Western blot assays. Solutions are activated by mixing two solutions in a 1:1 ratio for simple storage and activation.
Light Emission
LumiBlue ECL solutions use unique patented downstream catalysts and enhancers to greatly increase the brightness and stability of light produced by luminol and HRP. This allows flexible experimental design with increased time between blotting and analysis, as well as the ability to repeat exposures with negligible luminescence loss.
Compatibility with Standard Protocols
LumiBlue ECL kits have been developed to be compatible with standard ECL protocols used by other kits. In most cases experimental design does not need to be changed if switching between another ECL substrate and LumiBlue, but optimisation with LumiBlue ECL solutions may result in the use of more dilute antibodies for similar experiments.
LumiBlue ECL Range for Western Blot Detection
After establishing itself as a leader in protein staining with the InstantBlue single step 15 minute Coomassie stain, Expedeon has now launched its own range of ECL kits for protein detection in Western blotting. The LumiBlue ECL range consists of 5 kits with increasing levels of sensitivity to ensure reliable detection of protein ranging from high abundance expression to extremely dilute samples of protein and antibody. The LumiBlue ECL range has been designed to match existing kits to allow easy transition without protocol re-optimisation or changes to currently used methods. LumiBlue ECL solutions permit the use of less antibody while still obtaining excellent results, which often results in more economical use of precious antibodies.
**Human Transferrin serially diluted (5 to 0.5ng) and electrophoresis performed. The gels were transferred to PVDF membranes, blocked and incubated with different dilutions of rabbit anti-transferrin. After washing, the membranes were incubated with different dilutions of HRP-conjugated goat anti-rabbit antibody. The membranes were washed again and then incubated with substrate. Blots were acquired with ImageQuant LAS 4000 (GEHC) for 300 sec.
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