IRDye® 680RD Applications
Western Blot (see Fig. 1)
In-Cell Western™ Assay (see Fig. 2)
Figure 2. In-Cell Western™ Assay normalization with IRDye 680RD secondary antibody. A-431 cells were treated with 2-fold serial dilutions of EGF (800-1.6 ng/mL). After fixation and permeabilization, cells were probed with rabbit anti-phospho EGFR and Mouse anti-ERK2 followed by detection with IRDye 800CW Goat anti-Rabbit IgG (LI-COR P/N 926-32211) and IRDye 680RD Goat anti-Mouse IgG (LI-COR P/N 926-68070). Column 1=background (no primary antibody); Columns 2-11 = 2-fold dilution EGF treatment; Column 12 = 100% standard (no EGF treatment).
Figure 1. Western blot detection of HER2 in dose-response, Radicicol-treated SK-BR-3 cell lysate samples (treatment range 0.063 – 2.0 µM). The blot was probed with mouse anti-HER2 and rabbit anti-actin (LI-COR P/N 926-42210) followed by detection with IRDye 800CW Goat anti-Mouse IgG (LI-COR P/N 926-32210) and IRDye 680RD Goat anti-Rabbit IgG (LI-COR P/N 926-68071).