Quantitative chemiluminescent Western blotting
WesternLumaxLight Enhance is formulated to take full advantage of the linear dynamic range of CCD imagers.It provides high sensitivity
to detect low abundance proteins, and does not exhibit substrate depletionwith high abundance proteins, so has a larger linear dynamic
range than other commercially available substrates.A signal linear with respect to protein concentration over 3 orders of magnitude
provides theability to accurately quantify and compare protein amounts in your samples.
Figure 1. WesternLumaxLight Enhance 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. WesternLumaxLight Enhance 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. WesternLumaxLight Enhance produces the most stable chemiluminescent signal. Blots detected using WesternLumaxLight Enhance 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, WesternLumaxLight Enhance retains 70% of its initial signal strength, while the competition decays to 5% or less.
|
CUSTOMER FEEDBACK
We really like the Quantum kit. We use it instead of the ECL Plus kit because the signal lasts longer and doesn‘t burn out as frequently.
Assistant Research Professor,Ohio State University.
We love the Quantum. In our hands I’d say it’s 2-5x better than ECL Prime.
Associate Professor, University of Iowa Quantum gives alinear signal that is essential for protein quantification. Research Scientist, Ohio State University
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn