Western Blot Immunodetection of Cellular Proteins

Western blot was performed as described elsewhere [44 (link)]. The primary antibodies that were used were anti-P-glycoprotein (ab3366, RRID: AB_303744), anti-actin (ab11003, RRID: AB_297660) from Abcam (Cambridge, UK), and anti-TRIP6 (HPA052813, RRID: AB_2681961) from Atlas Antibodies (Bromma, Sweden). SuperSignal™ West Pico PLUS Chemiluminescent Substrate or West Femto Maximum Sensitivity Substrate (Pierce, Rockford, IL, USA) were applied on a membrane to detect the chemiluminescence signal of secondary HRP-conjugated goat anti-mouse (SA00001-1, RRID: AB_272565) and HRP-conjugated goat anti-rabbit (SA00001-2, RRID: AB_272564) from Proteintech (Rosemont, IL, USA). Images were obtained using ChemiDoc MP imaging system (Biorad, Hercules, CA, USA).

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Daniel P., Balušíková K., Václavíková R., Šeborová K., Ransdorfová Š., Valeriánová M., Wei L., Jelínek M., Tlapáková T., Fleischer T., Kristensen V.N., Souček P., Ojima I, & Kovář J. (2023). ABCB1 Amplicon Contains Cyclic AMP Response Element-Driven TRIP6 Gene in Taxane-Resistant MCF-7 Breast Cancer Sublines. Genes, 14(2), 296.

Publication 2023

ab3366 HRP-conjugated goat anti-rabbit (SA00001-2, ChemiDoc MP imaging system

Actin Antibodies Chemiluminescence Glycoprotein Goat Membrane Mouse Rabbit Sensitivity Western blot

Corresponding Organization : Charles University

Other organizations : National Institute of Public Health, Institute of Haematology and Blood Transfusion, Stony Brook University, State University of New York, Oslo University Hospital, University of Oslo

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Variable analysis

independent variables

Primary antibodies used: anti-P-glycoprotein, anti-actin, and anti-TRIP6

dependent variables

Chemiluminescence signal of secondary HRP-conjugated goat anti-mouse and HRP-conjugated goat anti-rabbit antibodies

control variables

Western blot protocol as described elsewhere [44]
SuperSignal™ West Pico PLUS Chemiluminescent Substrate or West Femto Maximum Sensitivity Substrate
ChemiDoc MP imaging system

controls

Positive control: None specified
Negative control: None specified

Annotations

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