Adherent cells were lysed by RIPA lysis buffer (CWBIO) containing protease inhibitors (CWBIO) on ice for 30 min. BCA kit (Thermo Fisher) was used for protein concentration determination. Detailed protocols of Western blot had been described previously.
34 (link) Primary antibodies in this study included anti‐DKK1 (1:1000, ab109416, Abcam), anti‐GAPDH (1:5000, ab8245, Abcam), anti‐pan‐Akt (1:1000, 4691S, Cell Signaling Technology), and anti‐Akt‐S473 (1:1000, 4060T, Cell Signaling Technology). The secondary antibodies included goat anti‐rabbit IgG H&L (HRP) (1:5000, ab6721, Abcam) and goat anti‐mouse IgG H&L (HRP) (1:5000, ab6789, Abcam). Images were captured by photo system (Mini Chemi 910, SinSage).
34 (link) Primary antibodies in this study included anti‐DKK1 (1:1000, ab109416, Abcam), anti‐GAPDH (1:5000, ab8245, Abcam), anti‐pan‐Akt (1:1000, 4691S, Cell Signaling Technology), and anti‐Akt‐S473 (1:1000, 4060T, Cell Signaling Technology). The secondary antibodies included goat anti‐rabbit IgG H&L (HRP) (1:5000, ab6721, Abcam) and goat anti‐mouse IgG H&L (HRP) (1:5000, ab6789, Abcam). Images were captured by photo system (Mini Chemi 910, SinSage).
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