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Afatinib bibw2992

Manufactured by Selleck Chemicals
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Afatinib (BIBW2992) is a small-molecule tyrosine kinase inhibitor. It is a chemical compound used as a laboratory reagent for research purposes.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Hek293t, by Thermo Fisher Scientific (1 mentions) Dmem f 12 l glutamine, by Thermo Fisher Scientific (1 mentions) Cetuximab erbitux, by Merck Group (1 mentions) Trametinib gsk1120212, by Selleck Chemicals (1 mentions) Mm hepes, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Trastuzumab, by Genentech (1 mentions) Phospho akt s473, by Cell Signaling Technology (1 mentions) Vegf c, by Abcam (1 mentions) Phospho faktyr925, by Cell Signaling Technology (1 mentions) Lyve 1, by Abcam (1 mentions) C myc, by Cell Signaling Technology (1 mentions) Stat3, by Cell Signaling Technology (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Lapatinib, by Chemietek (1 mentions) Recombinant epidermal growth factor egf, by Merck Group (1 mentions) Sp600125, by Selleck Chemicals (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Icr mice, by Daehan Biolink (1 mentions)

Close spelling variants of this product

BIBW2992 Afatinib

10 protocols using afatinib bibw2992

1

Cell Culture and Treatment with Targeted Therapies

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LIM1215, HCA7 and HEK-293T cell lines were cultured in DMEM/F-12 + L-Glutamine + 15 mM HEPES (Gibco) supplemented with 10% fetal bovine serum (FBS) (HyClone) and 1% penicillin/streptomycin (Gibco) at 37°C with 5% CO2. LIM1215 cells were obtained from the Ludwig Institute for Cancer Research, New York, NY, USA and HCA7 and HEK-293T cells were obtained from the American Type Culture Collection (ATCC).
Cetuximab (Erbitux®) was purchased from Merck Serono, lapatinib (Tykerb/Tyverb®) from GlaxoSmithKline, and trastuzumab (Herceptin®) and pertuzumab (Perjeta®) from Roche. Gefitinib (ZD1839), GDC-0941, trametinib (GSK1120212) and afatinib (BIBW2992) were purchased from Selleck Chemicals. NRG1 was purchased from Peprotech.
Bosch-Vilaró A., Jacobs B., Pomella V., Asbagh L.A., Kirkland R., Michel J., Singh S., Liu X., Kim P., Weitsman G., Barber P.R., Vojnovic B., Ng T, & Tejpar S. (2016). Feedback activation of HER3 attenuates response to EGFR inhibitors in colon cancer cells. Oncotarget, 8(3), 4277-4288.
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2

Combination Therapy for HER2-Positive Cancers

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Trastuzumab, a monoclonal antibody for HER2, was purchased from Genentech (South San Francisco, CA, USA). Dacomitinib (PF-00299804), an irreversible pan-HER inhibitor, and afatinib (BIBW-2992), a dual inhibitor of EGFR and HER2, were purchased from Selleck Chemicals LLC (Houston, TX, USA). Gemcitabine, cisplatin, and 5-fluorouracil (5-FU) were purchased from Lilly Korea Co., Seoul, Korea, JW Pharmaceutical Co., Seoul, Korea, and Ildong Pharmaceutical Co., Seoul, Korea, respectively.
Nam A.R., Kim J.W., Cha Y., Ha H., Park J.E., Bang J.H., Jin M.H., Lee K.H., Kim T.Y., Han S.W., Im S.A., Kim T.Y., Oh D.Y, & Bang Y.J. (2016). Therapeutic implication of HER2 in advanced biliary tract cancer. Oncotarget, 7(36), 58007-58021.
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3

Evaluating Afatinib's Molecular Mechanisms

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3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) (5 mg/mL; Sigma, St. Louis, MO, USA), dimethyl sulfoxide (DMSO; Sigma). Afatinib (BIBW2992) was purchased from Selleck Chemicals (Houston, TX). All these compounds were dissolved in DMSO at 10 mmol/L as stock solutions and stored at -20°C. Antibodies against GAPDH were from Epitomics (Burlingame, CA). Antibodies against CCR7 (Y59), VEGFR3, VEGFR2, VEGFC, and LYVE-1 were from Abcam (Cambridge, MA, USA). Antibodies against phospho-EGFR (Y1068), EGFR, Akt, phospho-Akt (S473), JAK1/2, phospho-JAK1 (Tyr1022/1023), phospho-JAK2 (Tyr1007/1008), c-Myc, STAT3, phospho-STAT3 (Tyr705), FAK, and phospho-FAK (Tyr925) were purchased from Cell Signaling Technology (Beverly, MA).
Zhang Y., Yang X., Liu H., Cai M, & Shentu Y. (2020). Inhibition of Tumor Lymphangiogenesis is an Important Part that EGFR-TKIs Play in the Treatment of NSCLC. Journal of Cancer, 11(1), 241-250.
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4

Kinase Inhibitors and Signaling Modulators

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Lapatinib was purchased from ChemieTek (Indianapolis, IN), afatinib (BIBW 2992) was from Selleckchem (Houston, TX), and canertinib (CI-1033, PD-183805) was kindly provided by Pfizer (Groton, CT), respectively. All kinase blockers were dissolved in DMSO and diluted 1:1,000 or 1:2,000 in medium before use. Recombinant epidermal growth factor (EGF) and heregulin-b1 (HRG-b1) were purchased from Sigma (St. Louis, MO). Fetal calf serum (FCS), DMEM, a-MEM, and RPMI 1640 were from GIBCO (Karlsruhe, Germany). The JNK inhibitor SP600125 and the STAT5 inhibitor II (IQDMA) were obtained from Selleckchem and from Merck (Darmstadt, Germany), respectively.
Gschwantler-Kaulich D., Grunt T.W., Muhr D., Wagner R., Kölbl H, & Singer C.F. (2016). HER Specific TKIs Exert Their Antineoplastic Effects on Breast Cancer Cell Lines through the Involvement of STAT5 and JNK. PLoS ONE, 11(1), e0146311.
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5

Afatinib and R-crizotinib Storage Protocol

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Afatinib (BIBW2992) and R-crizotinib (PF-02341066) were purchased from SelleckChem and were stored as per the manufacturer’s recommendation.
Das I., Wilhelm M., Höiom V., Franco Marquez R., Costa Svedman F., Hansson J., Tuominen R, & Egyhàzi Brage S. (2019). Combining ERBB family and MET inhibitors is an effective therapeutic strategy in cutaneous malignant melanoma independent of BRAF/NRAS mutation status. Cell Death & Disease, 10(9), 663.
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6

Osteoclast Differentiation in Mice

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Six-week-old male ICR mice were purchased from Dae Han Bio Link (Chungbuk, Korea). All animal experiments were approved by the committees on the care and use of animals in research at Kyungpook National University and were conducted in accordance with the guidelines for the care and use of laboratory animals. Recombinant mouse M-CSF and mouse RANKL were obtained from R&D Systems (Minneapolis, MN, USA). Afatinib (BIBW2992) was purchased from Selleckchem, (Houston, TX, USA). Fetal bovine serum (FBS) and α-minimum essential medium (α-MEM) were obtained from Gibco BRL (Grand Island, NY, USA).
Ihn H.J., Kim J.A., Bae Y.C., Shin H.I., Baek M.C, & Park E.K. (2017). Afatinib ameliorates osteoclast differentiation and function through downregulation of RANK signaling pathways. BMB Reports, 50(3), 150-155.
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7

Small Molecule Inhibitors in Cancer

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Afatinib (BIBW2992) and erlotinib (OSI-744) were purchased from Selleck Chemicals (TX, USA). The protein assay kit was from Bio-Rad (Hercules, CA, USA). The E-cadherin antibody was a gift from Dr. Keith R. Johnson (University of Nebraska Medical Center). All other antibodies used are listed in Supplementary Table 1.
Macha M.A., Rachagani S., Qazi A.K., Jahan R., Gupta S., Patel A., Seshacharyulu P., Lin C., Li S., Wang S., Verma V., Kishida S., Kishida M., Nakamura N., Kibe T., Lydiatt W.M., Smith R.B., Ganti A.K., Jones D.T., Batra S.K, & Jain M. (2017). Afatinib radiosensitizes head and neck squamous cell carcinoma cells by targeting cancer stem cells. Oncotarget, 8(13), 20961-20973.
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8

Antibody Acquisition and Characterization

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Afatinib (BIBW2992) and Erlotinib (OSI-420) were purchased from Selleck Chemicals (Houston, Texas, USA). Dimethyl sulfoxide (DMSO), pyrimethamine, and Sunitinib malate (SU 11248) were purchased from Sigma Aldrich (St. Louis, Missouri, USA). Bovine serum albumin was purchased from Bovogen Biologicals (Melbourne, Australia).
Antibodies against Phospho-STAT1 (Tyr701) (58D6), Phospho-STAT2 (Tyr690), Phospho-STAT3 (Ser727), Phospho-STAT5 (Tyr694) (C11C5), Phospho-STAT6 (Tyr641), and β-actin were purchased from Cell Signaling Technology (Beverly, Massachusetts, USA). Antibodies against STAT1 p91(c-111), STAT6 (M-20), Phospho-JAK1 (Tyr1022/Tyr1023), and Phospho-JAK3 (Tyr980) were purchased from Santa Cruz Biotechnology (Santa Cruz, California, USA). FITC-conjugated anti-mouse IgG antibody, TRTIC-conjugated anti-rabbit IgG antibody, and horseradish peroxidase-conjugated anti-rabbit or anti-mouse antibodies were purchased from Sigma Aldrich. Mouse Tg563 monoclonal antibody was cloned in our laboratory.
Yang Z., Ahn H.J., Park Y.H, & Nam H.W. (2016). Afatinib Reduces STAT6 Signaling of Host ARPE-19 Cells Infected with Toxoplasma gondii. The Korean Journal of Parasitology, 54(1), 31-38.
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9

Authentication and Validation of HaCaT Cell Line

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Cell line authenticity was confirmed using the Short Tandem Repeat (STR) Identifier kit (Applied Biosystems). HaCaT cells were cultured in W489 media consisting of 80% MCDB153 and 20% L15 medium supplemented with 1% FBS and maintained at 37°C in a humidified incubator with 5% CO2. All primary and secondary antibodies were purchased from Cell Signaling Technology (Boston, MA) except anti-HRAS antibody (Santa Cruz Biotechnology, Inc.). Cetuximab (Bristol-Myers Squibb, Princeton, NJ) was purchased from Johns Hopkins Pharmacy. Gefitinib was purchased from Tocris Bioscience (Ellisville, MO). Afatinib (BIBW2992) was purchased from Sellekchem. HaCaT, a spontaneously immortalized keratinocyte cell line was purchased from Cell Lines Service Germany. HaCaT cells were treated with Cetuximab (100nM) and Gefitinib (100nM), as described previously [10 (link), 62 (link)], and afatinib (10nM) based on the dose-response curves for HaCaT-Mock cells. HNSCC cells are grown and treated with Cetuximab (100 nM), as previously described [10 (link), 62 (link)].
Fertig E.J., Ozawa H., Thakar M., Howard J.D., Kagohara L.T., Krigsfeld G., Ranaweera R.S., Hughes R.M., Perez J., Jones S., Favorov A.V., Carey J., Stein-O'Brien G., Gaykalova D.A., Ochs M.F, & Chung C.H. (2016). CoGAPS matrix factorization algorithm identifies transcriptional changes in AP-2alpha target genes in feedback from therapeutic inhibition of the EGFR network. Oncotarget, 7(45), 73845-73864.
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10

Afatinib Cytotoxicity Evaluation in Vitro

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Afatinib (BIBW2992) was purchased from Selleck Chemicals LLC (USA); fetal bovine serum (FBS), phosphate buffered saline (PBS), trypsin, Dulbecco's modified Eagle's medium (DMEM) and RPMI 1640 medium were obtained from Invitrogen Co. (Scotland, UK). Acetic acid, dimethyl sulfoxide (DMSO), sulforhodamine B (SRB), trypan blue, trisodium citrate dehydrate, tetrachloroauric (III) acid (HAuCl4; 99.99% trace metals basis, 30 wt % in dilute HCl, N-ethyl-N'-(3-dimethylaminopropyl) cabodiimide (EDC), Sulfo-NHS (N-hydroxysulfosuccinimide) and propidium iodide were purchased from Sigma-Aldrich (Germany).
TricloroAcetic acid (TCA) and Tris buffer were acquired from Merck (Darmstadt, Germany). a-thiol-wcarboxyl (polyethylene glycol) (HS-PEG-COOH; molecular weight 394.57 Da) was purchased from Prochimia (Poland).
Coelho S.C., Almeida G.M., Pereira M.C., Santos-Silva F, & Coelho M.A. (2016). Functionalized gold nanoparticles improve afatinib delivery into cancer cells. Expert opinion on drug delivery, 13(1).
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