- Sensitive – detect attomoles of protein per band.
- Quantitative – linear range of signal with respect to protein amount exceeds 3 orders of magnitude
- Low background – for high signal to noise
- Long lasting signal – image blots hours after substrate incubation
- Versatile – Optimized for CCD imaging, and compatible with film detection. In addition, chemifluorescent emissions can be detected with fluorescence imaging systems
WesternBright Quantum chemiluminescent substrate sets the bar for both sensitivity and quantitative ability. Specially developed for CCD imaging, WesternBright Quantum produces a strong, long-lasting signal with extremely low background, perfect for detecting low abundance proteins. Since it does not exhibit substrate depletion at high protein loads, WesternBright Quantum provides the largest dynamic range of any chemiluminescent substrate for the most quantitative chemiluminescent Western experiments.
Figure 1. WesternBright Quantum provides a larger linear dynamic range than Amersham ECL or ECL Plus (GE Healthcare) or SuperSignal West Dura (Thermo Scientific). Identical Western blots containing serial dilutions of transferring were probed with a rabbit-anti-transferrin primary antibody and a goat-anti-rabbit secondary antibody conjugated to horseradish peroxidase. The blots were incubated with chemiluminescent substrates as recommended by each manufacturer. All blots were imaged simultaneously for 2 minutes on a CCD imager and display parameters are identical across all images shown. Band intensities were plotted and a best fit linear regression conducted for each substrate. WesternBright Quantum shows the largest linear dynamic range out of all four substrates with the highest R2 value. Bands that fall on the linear part of the curve for each substrate are indicated in the image.
Figure 2. WesternBright Quantum produces the most stable chemiluminescent signal. Blots detected using WesternBright Quantum or one of three other chemiluminescent substrates were re-imaged at several times over a 10 hour period. The intensitiy of one band is plotted. 60 minutes after substrate incubation, WesternBright Quantum retains 70% of its initial signal strength, while the competition decays to 5% or less.