Egfr inhibitor ag1478

Manufactured by Merck Group

Sourced in United States

EGFR inhibitor (AG1478) is a laboratory research tool compound that inhibits the epidermal growth factor receptor (EGFR) protein. It is commonly used in experimental studies to investigate the role of EGFR signaling in various cellular processes.

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4 protocols using egfr inhibitor ag1478

Molecular Mechanisms of Cell Signaling Regulation

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A rabbit polyclonal anti-actin antibody, EGFR inhibitor (AG1478) and 2-deoxy-D-glucose (2-DG) were obtained from Sigma-Aldrich (St. Louis, Mo., USA). Dobutamine hydrochloride was obtained from Wako Laboratory Chemicals (Tokyo, Japan). Recombinant human amphiregulin was obtained from PeproTech (Rocky Hill, NJ). Rabbit polyclonal anti-lipolysis-stimulated receptor (LSR) and anti-tricellulin (TRIC) antibodies were obtained from Zymed Laboratories (San Francisco, CA). A mouse monoclonal anti-LSR antibody was obtained from Abnova (Yaipei, Taiwan). A rabbit polyclonal anti-angiomotin (AMOT) antibody was obtained from Bioss, Inc. (Woburn, MA). Rabbit monoclonal anti-Merlin, anti-phospho- YAP (ser127) and anti-phospho-AMPK (ther172) antibodies were obtained from Cell Signaling Technology Japan (Tokyo, Japan). A rabbit polyclonal anti-YAP antibody was obtained from Novus Biologicals (Littleton, CO). Alexa 488 (green)-conjugated anti-rabbit IgG and Alexa 594 (red)-conjugated anti-mouse IgG antibodies were purchased from Molecular Probes, Inc. (Eugene, OR).

Shimada H., Abe S., Kohno T., Satohisa S., Konno T., Takahashi S., Hatakeyama T., Arimoto C., Kakuki T., Kaneko Y., Takano K.I., Saito T, & Kojima T. (2017). Loss of tricellular tight junction protein LSR promotes cell invasion and migration via upregulation of TEAD1/AREG in human endometrial cancer. Scientific Reports, 7, 37049.

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Inhibition of MAPK Signaling Pathways

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suPAR and ATF were purchased from American Diagnostica, Inc. (Greenwich, CT). fMLP, Pertussis toxin (Gαi inhibitor, 100ng/ml), Phosphatidylinositol-specific-phospholipase C (100µg/mL), PD98059 (ERK inhibitor, 25µM) SB203580 (p38^MAPK inhibitor, 10µM), SP600125 (JNK inhibitor, 1µM), EGFR inhibitor ( AG1478, 10µM), ε-aminocaproic acid (EACA; plasmin inhibitor, 100µM), aprotinin (plasmin inhibitor, 100 units/ml) and GM6001 (MMP inhibitor, 10nM) were purchased from Sigma Chemical Co. (St. Louis, MO). Peroxidase-conjugated anti-rabbit IgG antibody (raised in goat) and peroxidase-conjugated anti-mouse IgG antibody (raised in goat) were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA). Phospho-ERK1/2 antibody was purchased from Promega, Inc. (Madison, WI). Total ERK 1/2 antibody was purchased from BD Transduction Laboratories (Lexington, KY). Phospho-p38^MAPK and phospho-JNK antibodies were purchased from Biosource (Camarillo, CA). Total p38^MAPK and JNK antibodies were purchased from Cell Signaling (Beverley, MA). Dulbecco’s minimal essential medium (DMEM) and Dulbecco’s phosphate buffered saline (dPBS) were purchased from Corning Cellgro (Corning, NY).

Duru E.A., Fu Y, & Davies M.G. (2015). ROLE OF FORMIC RECEPTORS IN SOLUBLE UROKINASE RECEPTOR INDUCED HUMAN VASCULAR SMOOTH MUSCLE MIGRATION. The Journal of surgical research, 195(2), 396-405.

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Growth Factor Activation Pathway Protocol

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EGF was purchased from Invitrogen, Carlsbad, CA, USA. AREG and TGF-

α

were from R&D Systems, Minneapolis, MN, USA. If not otherwise indicated, 10 nM of EGF and 20 nM AREG or TGF-

α

were used. EGFR inhibitor AG-1478 (Sigma-Aldrich, Saint Louis, MO, USA) was used at a concentration of 1

μ

M. The protein synthesis inhibitor cycloheximide (CHX, Sigma-Aldrich, USA) was used at a concentration of 35.5

μ

M. The vacuolar V-ATPase inhibitor bafilomycin A

_{1}

(BafA, Sigma-Aldrich, USA) was used at a concentration of 100 nM.

Kamentseva R.S., Kharchenko M.V., Gabdrahmanova G.V., Kotov M.A., Kosheverova V.V, & Kornilova E.S. (2023). EGF, TGF-α and Amphiregulin Differently Regulate Endometrium-Derived Mesenchymal Stromal/Stem Cells. International Journal of Molecular Sciences, 24(17), 13408.

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Antibodies for Tight Junction Analysis

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Rabbit polyclonal anti-tricellulin (TRIC), anti-claudin (CLDN)-1 and -2 antibodies and mouse monoclonal CLDN-2 and anti-occludin (OCLN) antibodies were obtained from Zymed Laboratories (San Francisco, CA, USA). Rabbit polyclonal anti-lipolysis-stimulated lipoprotein receptor (LSR) antibodies were from Novus Biologicals (Littleton, CO, USA). A mouse monoclonal anti-LSR antibody was obtained from Abnova (Taipei, Taiwan). A rabbit polyclonal anti-p63 antibody was obtained from Abcam (Cambridge, UK). Mouse monoclonal anti-acetylated tubulin and antiCK7 antibodies and a rabbit polyclonal anti-actin antibody were from Sigma-Aldrich (St. Louis, MO, USA). Alexa 488 (green)-conjugated anti-rabbit IgG and Alexa 594 (red)-conjugated anti-mouse IgG antibodies and Alexa 594 (red)-conjugated phalloidin were from Molecular Probes, Inc. (Eugene, OR, USA). HRP-conjugated polyclonal goat anti-rabbit IgG was from Dako A/S (Glostrup, Denmark). The ECL Western blotting system was from GE Healthcare UK, Ltd. (Buckinghamshire, UK). A rabbit polyclonal anti-actin antibody and EGFR inhibitor (AG1478) were obtained from Sigma-Aldrich (St. Louis, MO, USA). A TGF-β receptor type 1 inhibitor (EW-7197) was obtained from Cayman Chemical (Ann Arbor, MI, USA). FITC-dextran (FD-4, MW 4.0 kDa) was obtained from Sigma-Aldrich Co. (St. Louis, MO, USA).

Arai W., Konno T., Kohno T., Kodera Y., Tsujiwaki M., Shindo Y., Chiba H., Miyajima M., Sakuma Y., Watanabe A, & Kojima T. (2023). Downregulation of angulin-1/LSR induces malignancy via upregulation of EGF-dependent claudin-2 and TGF-β-dependent cell metabolism in human lung adenocarcinoma A549 cells. Oncotarget, 14, 261-275.

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