Advansta
WesternBright ECL HRP substrate
K-12045
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WesternBright ECL HRP substrate from Advansta was used to detect by western blotting a specific protein in comparison with Thermo Scientific West Pico reagent used conventionally in our lab.
Western blotting
Cell lysates; western blot equipment and reagents to run the SDS gels and transfer separated proteins onto a PVDF membrane.
WesternBright ECL HRP substrate was used in comparison with West Pico as recommended by the manufacturers. WesternBright ECL detected the similar amount of specific protein run in the gel (e.g., 20 ug per lane), but the development of blot required 5 times less primary antibody (0.2 ug/ml vs. 1 ug/ml
If the amount of a protein of interest is not limited, do not reduce too much the total amount of protein to be separated by SDS-PAGE. It is better to use less primary and secondary antibody.
WesternBright ECL detected the similar amount of specific protein run in the gel (e.g., 20 ug per lane), but development of a blot required 5 times less primary antibody (0.2 ug/ml vs. 1 ug/ml) and 3.3 times less secondary antibody (1:10,000 vs 1:3,000). WesternBright ECL also detected with higher efficiency mol. wt. markers.
N/A
The image depicts detection by "Advansta" or "West Pico" of 3 specific bands of a protein (20 ug total protein per lane).
Advansta produces great results in western blotting.
None detected
WesternBright ECL HRP substrate from Advansta is a great reagent to use in western blotting especially when specific proteins or primary antibodies are limited.