Afatinib

Manufactured by LC Laboratories

Sourced in United States

Afatinib is a small-molecule tyrosine kinase inhibitor that targets the epidermal growth factor receptor (EGFR) family, including EGFR (ErbB1), HER2 (ErbB2), ErbB3, and ErbB4. It is used for research purposes in various biological and biochemical applications.

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Lab products found in correlation

Erlotinib, by LC Laboratories (13 mentions) Lapatinib, by LC Laboratories (5 mentions) Osimertinib, by Selleck Chemicals (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Neratinib, by LC Laboratories (3 mentions) Crizotinib, by LC Laboratories (3 mentions) Osimertinib, by LC Laboratories (3 mentions) Rpmi 1640 growth medium, by Thermo Fisher Scientific (3 mentions) Cetuximab, by Merck Group (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Phospho egfr y1068 antibody 44788g, by Thermo Fisher Scientific (2 mentions) Linsitinib, by LC Laboratories (2 mentions) Imatinib, by LC Laboratories (2 mentions) Dacomitinib, by Selleck Chemicals (2 mentions) Selumetinib, by Selleck Chemicals (2 mentions) Mobocertinib, by MedChemExpress (2 mentions) Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions)

35 protocols using afatinib

Afatinib Attenuates OGD-Induced Astrocyte Activation

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Afatinib was purchased from LC laboratories, MA, U.S.A. Afatinib (10 mM) was prepared in dimethyl sulfoxide as stock solution. The effect of Afatinib was investigated by treating Afatinib concomitantly with OGD exposure. Ten nM Afatinib was chosen according to the concentration of Afatinib which significantly attenuated OGD-induced activation of EGFR and subsequent singalling pathways in treated astrocytes.

Chen Y.J., Hsu C.C., Shiao Y.J., Wang H.T., Lo Y.L, & Lin A.M. (2019). Anti-inflammatory effect of afatinib (an EGFR-TKI) on OGD-induced neuroinflammation. Scientific Reports, 9, 2516.

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Lapatinib and Trastuzumab Stock Preparation

Cited 2 times

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Lapatinib and trastuzumab were purchased from LC Laboratories and Mckesson Specialty Health, respectively. Stocks of L and T were prepared as described previously (10 (link), 19 (link)). Afatinib (Afa) and neratinib (Nrb) were purchased from LC Laboratories and Selleck Chemicals, respectively. Stocks of Afa and Nrb were prepared with DMSO.

Xu X., De Angelis C., Burke K.A., Nardone A., Hu H., Qin L., Veeraraghavan J., Sethunath V., Heiser L.M., Wang N., Ng C.K., Chen E.S., Renwick A., Wang T., Nanda S., Shea M., Mitchell T., Rajendran M., Waters I., Zabransky D.J., Scott K.L., Gutierrez C., Nagi C., Geyer F.C., Chamness G.C., Park B.H., Shaw C.A., Hilsenbeck S.G., Rimawi M.F., Gray J.W., Weigelt B., Reis-Filho J.S., Osborne C.K, & Schiff R. (2017). HER2 Reactivation through Acquisition of the HER2 L755S Mutation as a Mechanism of Acquired Resistance to HER2-targeted Therapy in HER2+ Breast Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(17), 5123-5134.

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Comparative Analysis of EGFR Inhibitors

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Erlotinib and afatinib were purchased from LC Laboratories (Woburn, MA, USA). Osimertinib and rociletinib were purchased from Selleck Chemicals (Houston, TX, USA). Total EGFR antibody (#2232), total AKT antibody (#9272), phospho-AKT (S473; D9E) antibody (#4060), total p44/42 MAPK antibody (#9102S), and phospho-p44/42 MAPK (T202/204) antibody (#9101S) were purchased from Cell Signaling Technology (Beverly, MA, USA). Phospho-EGFR (Y1068) antibody (44788G) was purchased from Invitrogen/Life Technologies (Carlsbad, CA, USA). Actin antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA).

Hirano T., Yasuda H., Tani T., Hamamoto J., Oashi A., Ishioka K., Arai D., Nukaga S., Miyawaki M., Kawada I., Naoki K., Costa D.B., Kobayashi S.S., Betsuyaku T, & Soejima K. (2015). In vitro modeling to determine mutation specificity of EGFR tyrosine kinase inhibitors against clinically relevant EGFR mutants in non-small-cell lung cancer. Oncotarget, 6(36), 38789-38803.

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Composition and Characterization of HAD-B1 Herbal Extract

Cited 1 time

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Afatinib was purchased from LC laboratories (Woburn, MA, USA). HAD-B1 is a
blended herbal extract composed of Panax notoginseng Radix,
Cordyceps militaris, Panax ginseng C. A.
Mey, and Boswellia carteri Birdwood (Table 1). It was manufactured by
Daehan Biopharm (Gyeonggi, Republic of Korea) and provided by the EWCC (Daejeon
Korean Medicine Hospital, Daejeon University, Korea) in August 2019. In this
study, HAD-B1 suspended in dimethyl sulfoxide was centrifuged, and only the
supernatant was used.
A previous study identified 6 critical components of HAD-B1 by using high
performance liquid chromatography (HPLC).^{24 (link)} Each HAD-B1 tablet
contained 0.320 mg of cordycepin, 1.110 mg of notoginsenoside R1, 0.610 mg of
ginsenoside Rg1, 1.270 mg of ginsenoside Rb1, 0.031 mg of α-boswellic acid, and
0.058 mg of β-boswellic acid.

Song S.Y., Park J.H., Park S.J., Kang I.C, & Yoo H.S. (2022). Synergistic Effect of HAD-B1 and Afatinib Against Gefitinib Resistance of Non-Small Cell Lung Cancer. Integrative Cancer Therapies, 21, 15347354221144311.

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Tyrosine Kinase Inhibitor Dissolution

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Erlotinib, afatinib and rociletinib were purchased from LC Laboratories. Luminespib was purchased from Selleckchem. All reagents were dissolved in dimethyl sulfoxide and stored at −80°C.

Jorge S.E., Lucena-Araujo A.R., Yasuda H., Piotrowska Z., Oxnard G.R., Rangachari D., Huberman M.S., Sequist L.V., Kobayashi S.S, & Costa D.B. (2018). EGFR exon 20 insertion mutations display sensitivity to Hsp90 inhibition in preclinical models and lung adenocarcinomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(24), 6548-6555.

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Small Molecule Dissolution and Storage

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Crizotinib and afatinib were purchased from LC Laboratories (Woburn, MA). Ceritinib was purchased from Active Biochemicals (Hong Kong). All reagents were dissolved in dimethyl sulfoxide (DMSO) and stored at −80°C.

Lucena-Araujo A.R., Moran J.P., VanderLaan P.A., Dias-Santagata D., Folch E., Majid A., Kent M.S., Gangadharan S.P., Rangachari D., Huberman M.S., Kobayashi S.S, & Costa D.B. (2016). De novo ALK kinase domain mutations are uncommon in kinase inhibitor-naïve ALK rearranged lung cancers. Lung cancer (Amsterdam, Netherlands), 99, 17-22.

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Cell Lines and Inhibitor Characterization

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Cell lines, inhibitors and growth factors. The cell lines used in the present study included ALK translocated NSCLC lines H3122 and H2228, and their counterparts modified to overexpress GFP or HER2 (H3122) or to knock down GFP or HER2 (H2228). The original cell lines were a kind gift from Dr Pasi Jänne (Dana-Farber Cancer Institute, Boston, MA, USA). The cell lines were grown in RPMI-1640 medium (Thermo Fisher Scientific, Waltham, MA, USA and Sigma-Aldrich, St. Louis, MO, USA) with 10% fetal bovine serum (FBS) and 100 IU/ml penicillin and streptomycin (Thermo Fisher Scientific). Cells were incubated at 37˚C with 5% CO 2 in the atmosphere. Following inhibitors were used: TAE684 (a gift from Dr Nathanael Gray, Dana-Farber Cancer Institute), crizotinib, afatinib (LC Laboratories, Woburn, MA, USA) and labatinib (Alexis Biochemicals, Lausen, Switzerland). Neuregulin-1 (NRG1) and epidermal growth factor (EGF) were also used (Thermo Fisher Scientific). Inhibitors were diluted in dimethyl sulfoxide (DMSO) and stored at -20˚C while growth factors were diluted in sterile, distilled water and stored at -80˚C.

Honkanen T., Wilenius E., Koivunen P, & Koivunen J.P. (2017). HER2 regulates cancer stem-like cell phenotype in ALK translocated NSCLC. International journal of oncology, 51(2).

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Cell Line Maintenance and Drug Acquisition

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H1975 cells harboring EGFR L858R and T790M mutation were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). PC9 cells (EGFR exon 19del) were obtained from Dr. Joan Massague (Memorial Sloan Kettering Cancer Center, New York, NY) and H3255 cells (EGFR L858R) were obtained from NCI (National Cancer Institute, Bethesda, MD). PC9, H3255, H1975, and AR (afatinib resistant) cells were maintained in RPMI-1640 growth medium supplemented with 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA) in a 37°C humidified 5% CO2 incubator. afatinib and linsitinib was purchased from LC laboratories (Massachusetts, USA).

Lee Y., Wang Y., James M., Jeong J.H, & You M. (2015). Inhibition of IGF1R Signaling Abrogates Resistance to Afatinib (BIBW2992) in EGFR T790M Mutant Lung Cancer Cells. Molecular carcinogenesis, 55(5), 991-1001.

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Evaluating ERBB2-Mutant Cell Response to TKIs

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NCI-H1781 (H1781) cells, harboring ERBB2-G776>VC, were purchased from ATCC (Manassas, VA). Cells were maintained in RPMI 1640 medium (Mediatech, Manassas, VA) supplemented with 10% fetal bovine serum and grown at 37°C in a humidified atmosphere with 5% CO₂. These cells were overlaid in 96-well plates, allowed to attach overnight and then treated with or without two TKIs (Erlotinib and afatinib) for 72 hours. Cell viability was determined by CellTiter 96 Aqueous One solution proliferation kit (Promega, Madison, WI) according to the manufacture's protocol. Experiments were performed in triplicate. Inhibitory proliferation curves and the 50% inhibitory concentration (IC₅₀) were generated using the GraphPad Prism 6 software (GraphPad Software, La Jolla, CA). We used IC₅₀ values obtained in identical conditions and previously published [11 ] for NCI-H3255 (H3255, EGFR-L858R) and NCI-H1975 (H1975, EGFR-L858R+T790M) to contrast H1781 to an EGFR/ERBB2 TKI-sensitive and TKI-resistant cell line, respectively. Erlotinib and afatinib were purchased from LC Laboratories (Woburn, MA). All reagents were dissolved in dimethyl sulfoxide and stored at −80°C.

Costa D.B., Jorge S.E., Moran J.P., Freed J.A., Zerillo J.A., Huberman M.S, & Kobayashi S.S. (2016). Pulse afatinib for ERBB2 exon 20 insertion mutated lung adenocarcinomas. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 11(6), 918-923.

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Gefitinib-sensitive and resistant NSCLC cell lines

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The gefitinib-sensitive human adenocarcinoma NSCLC cell line PC9 (EGFR exon19del E746-A750) was kindly provided by Dr Pan-Chyr Yang, and gefitinib-resistant NSCLC H1975 cells (EGFR L858R/T790M; IC₅₀ >10 μM) were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA). All cells were maintained in RPMI 1640 growth medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (Thermo Fisher Scientific), penicillin, and streptomycin (Thermo Fisher Scientific) in humidified 5% CO₂ at 37°C. EGFR TKIs gefitinib (Ryss Lab, Inc., Union City, CA, USA), afatinib (LC Laboratories, Woburn, MA, USA), Src TKI dasatinib (LC Laboratories), and integrin inhibitor cilengitide (ci) (AdooQ Bioscience, Irvine, CA, USA) were obtained from commercial sources. The integrin inhibitor c8 was kindly provided by Dr William F DeGrado.9 Stock solutions (10 mM) of all chemicals were prepared in dimethyl sulfoxide (DMSO). Both cell lines used in the current study can be obtained commercially and they were classified as the most low risk by the institutional review board of National Health Research Institute. The ethics approval was not required for the use of these cell lines.

Wang H.Y., Hsu M.K., Wang K.H., Tseng C.P., Chen F.C, & Hsu J.T. (2016). Non-small-cell lung cancer cells combat epidermal growth factor receptor tyrosine kinase inhibition through immediate adhesion-related responses. OncoTargets and therapy, 9, 2961-2973.

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