GenScript's One-Step WesternTM Fluorescent Kit yields journal quality Western or Dot blot results in about one hour. Using GenScript’s breakthrough immunodetection technology (patent pending), the kit replaces the classical three-step Western process which takes nearly five hours. Transfer the proteins from gel to membrane and incubate it in the pretreat solution for five minutes. Then incubate the membrane in WB solution with primary antibody and secondary antibody labeled with fluorescent dye for 40 minutes. After washing three times for ten minutes each, the membrane can then be scanned on LI-COR Odyssey Infrared Imaging System.
Key Features:
One-Step WesternTM Kits cut Western blot analysis time down to ONE hour.
Any primary antibody from any species can be used.
More sensitive than traditional chemiluminescence methods.
Examples:
Fluorescent Western blot detection of GAPDH using Goat Anti-GAPDH Polyclonal Antibody (GenScript, A00191).
Figure 1. Fluorescent Western blots for the detection of GAPDH using the One-Step WesternTM Fluorescent Kit (GenScript, L00397). 5.0, 2.5, 1.25, 0.62, 0.31, 0.16, 0.08 and 0.04 µg of HeLa cell lysates were loaded into lanes 1, 2, 3, 4, 5, 6, 7 and 8 respectively.
Mixture 1 was prepared by mixing 6 µg of Goat Anti-GAPDH Polyclonal Antibody (GenScript, A00191) with 3 µg of IRDye®800CW Donkey anti-Goat (LI-COR) in 100 µl of WB-1.
