Western Blot Analysis of P-gp Protein

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)].

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Wu C.P., Hung C.Y., Murakami M., Wu Y.S., Lin C.L., Huang Y.H., Hung T.H., Yu J.S, & Ambudkar S.V. (2022). P-glycoprotein Mediates Resistance to the Anaplastic Lymphoma Kinase Inhiitor Ensartinib in Cancer Cells. Cancers, 14(9), 2341.

Publication 2022

anti-alpha-tubulin Horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G (IgG)

Alpha tubulin Antibodies Antibody Dilution Goat Horseradish peroxidase Immunoglobulin g Mouse Protein Tubulin Western blot analysis

Corresponding Organization : Chang Gung Memorial Hospital

Other organizations : National Cancer Institute, Center for Cancer Research, Tunghai University, Keelung Chang Gung Memorial Hospital, Linkou Chang Gung Memorial Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression of P-gp protein

control variables

Dilution of primary antibody for P-gp (1:3000)
Dilution of primary antibody for alpha-tubulin (1:100,000)
Dilution of secondary antibody (1:100,000)
Experimental procedure for Western blot analysis as described previously [41]
Signals detected as previously described [41]

positive controls

Alpha-tubulin as a positive loading control

negative controls

None explicitly mentioned

Annotations

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