Horseradish peroxidase conjugated goat anti mouse immunoglobulin g igg

Western blot analysis of P-gp protein was performed as described previously [41 (link)]. The primary antibodies C219 at 1:3000 (#517310, Merck Millipore, Burlington, MA, USA), and anti-alpha-tubulin at 1:100,000 dilution (#T6199, Sigma-Aldrich, St. Louis, MO, USA) were used to identify P-gp, with tubulin as the positive loading control. Horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G (IgG) 1:100000 dilution (Abcam, Cambridge, MA, USA) was used as the secondary antibody. Signals were detected as previously described [41 (link)].

Immunoblotting was carried out following the standard procedure as previously described [20 (link)]. In brief, cancer cells were treated with DMSO (control) or furmonertinib at concentrations of 20 nM, 100 nM, 200 nM, or 1000 nM for 72 h. After the treatment, the cells were harvested and subjected to SDS-polyacrylamide electrophoresis (SDS-PAGE). For Western blotting, the primary antibody C219 (diluted 1:3000, Merck Millipore, Burlington, MA, USA) was used to detect human ABCB1, the primary antibody BXP-21 (diluted 1:15,000, Abcam, Cambridge, MA, USA) was used to detect human ABCG2, and the primary antibody anti-alpha tubulin (diluted 1:100,000, Sigma-Aldrich, St. Louis, MO, USA) was used as a positive loading control for tubulin. A horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G (IgG) (diluted 1:100,000, Abcam, Cambridge, MA, USA) was used as the secondary antibody. Signal detection was performed using the enhanced chemiluminescence (ECL) kit (Merck Millipore, Billerica, MA, USA), as described in previous work [20 (link)].