Western Blot Analysis of Cellular Proteins

Western blot was performed as previously described by our group (31 (link)) using the following primary antibodies: VIM (rabbit polyclonal, dilution 1:3,000, catalog number 10366-1-AP, ProteinTech, Manchester, UK), CDH1 (mouse monoclonal, Clone 36, dilution 1:500, catalog number 610182, BD Transduction Laboratories, San Jose, CA, USA) and α-actin (mouse monoclonal, Clone C4, dilution 1:1,000, catalog number 691001, MP Biomedicals, Solon, OH, USA). Antibody binding was revealed by incubation with anti-mouse (rabbit polyclonal, dilution 1:5,000, catalog number A9044, Sigma-Aldrich, Merk Merck Life Science, Madrid, Spain) or anti-rabbit (goat polyclonal, dilution 1:5,000, catalog number sc-2004, Santa Cruz Biotechnology, Dallas, TX, USA) IgG peroxidase conjugate secondary antibodies. Protein expression was assessed using HPR Chemiluminescent Substrate Reagent Kit (catalog number WP20005, Invitrogen, Thermo Fisher Scientific) and ChemiDoc (BIO-RAD, Hercules, CA, USA).

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Martín J., Castellano J.J., Marrades R.M., Canals J., Viñolas N., Díaz T., Molins L., Martinez D., Han B., Moisés J., He Y., Monzó M, & Navarro A. (2021). Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer. Translational Lung Cancer Research, 10(4), 1804-1818.

Publication 2021

α-actin ChemiDoc

Actin Antibodies Antibody Cdh1 Clone Dilution Goat Mouse Peroxidase Protein Rabbit Solon Western blot

Corresponding Organization : Hospital Clínic de Barcelona

Other organizations : Centro de Investigación Biomédica en Red de Enfermedades Respiratorias

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Variable analysis

independent variables

Primary antibodies used: VIM (rabbit polyclonal, dilution 1:3,000), CDH1 (mouse monoclonal, Clone 36, dilution 1:500), and α-actin (mouse monoclonal, Clone C4, dilution 1:1,000)

dependent variables

Protein expression assessed using HPR Chemiluminescent Substrate Reagent Kit and ChemiDoc

control variables

Western blot protocol as previously described by the group (31)
Anti-mouse (rabbit polyclonal, dilution 1:5,000) and anti-rabbit (goat polyclonal, dilution 1:5,000) IgG peroxidase conjugate secondary antibodies

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