Nilotinib

Manufactured by Selleck Chemicals

Sourced in United States, Germany, France

Nilotinib is a synthetic chemical compound that functions as a tyrosine kinase inhibitor. It is primarily used in research and laboratory settings to study cellular signaling pathways and biochemical processes.

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Lab products found in correlation

53 protocols using nilotinib

Inhibition of Stat1 and DDR1 in CAM Assay

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Fludarabine (Stat 1 inhibitor) was purchased from Selleckchem (reference S1491), diluted in DMSO at 50mM and used for 24hrs at a final concentration of 20μM.
Cells were pretreated overnight before inoculation in chicken fertilized eggs with Fludarabine 20μM. Control cells were treated with similar DMSO concentration. Tumors were then processed as described in CAM assay section of this article.
Nilotinib (DDR1 inhibitor) was purchased from Selleckchem (reference AMN-107) diluted in DMSO at 2.5mM and used 24hrs at a final concentration of 10μM. Cells were pretreated overnight before inoculation in chicken fertilized eggs with Nilotinib 10μM. Control cells were treated with similar DMSO concentration. Tumors were then processed as described in CAM assay section of this article.

Di Martino J.S., Nobre A.R., Mondal C., Taha I., Farias E.F., Fertig E.J., Naba A., Aguirre-Ghiso J.A, & Bravo-Cordero J.J. (2021). A tumor-derived type III collagen-rich ECM niche regulates tumor cell dormancy. Nature cancer, 3(1), 90-107.

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Inhibition of Stat1 and DDR1 in CAM Assay

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High-Throughput Screening of Tyrosine Kinase Inhibitors

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HGF: 20 ng/ml, EGF: 30 ng/ml, bFGF: 100 ng/ml (all from Preprotech). Drugs: (all from Selleckchem, Pittsburg, PA, USA): SCH772984: 1 µM, Staurosporin: 1 µM, H₂O₂: 800 mM. Axitinib: 1 µM, 5 µM, 10 µM, NPV-BEZ35: 250 nM, 500 nM, 1 µM, 5 µM, 10 µM, dasatinib (BMS-354825): 250 nM, 1 µM, 5 µM, 10 µM, Erlotinib HCI: 250 nM, 500 nM, 1 µM, 5 µM, 10 µM, Imatinib mesylate: 1 µM, 5 µM, 10 µM, Nilotinib (AMN-107): 1 µM, 5 µM, 10 µM, Rapamycin (sirolimus): 250 nM, 500 nM, 1 µM, 5 µM, 10 µM, Sunitinib malate (sutent): 1 µM, 5 µM, 10 µM, Temsirolimus (torisel): 1 µM, 5 µM, 10 µM, Masitinib (AB1010): 500 nM, 1 µM, 5 µM, 10 µM, Crizotinib (PF-02341066): 250 nM, 500 nM, 1 µM, 5 µM, 10 µM, Foretinib: 500 nM, 1 µM, 5 µM, 10 µM (Selleckchem), XL-184: 1 µM, 5 µM, 10 µM, Vermurafenib (PLX4032): 1 µM, 5 µM, 10 µM, Zoledronic acid (zoledronate): 1 µM, 5 µM, 10 µM, Ruxolitinib (INCB018424): 500 nM, 1 µM, 5 µM, 10 µM, Nystatin (mycostatin): 1 µM, 5 µM, 10 µM, lapatinib: 1 µM, 5 µM, 10 µM, Pazopanib: 1 µM, 5 µM, 10 µM, dequalinium chloride: 1 µM, 5 µM, 10 µM, Tofacitinib citrate (CP-690550 citrate): 500 nM, 1 µM, 5 µM, 10 µM. H-1152 (from Alexis Biochemicals): 1 µM.

Schönholzer M.T., Migliavacca J., Alvarez E., Santhana Kumar K., Neve A., Gries A., Ma M., Grotzer M.A, & Baumgartner M. (2020). Real-time sensing of MAPK signaling in medulloblastoma cells reveals cellular evasion mechanism counteracting dasatinib blockade of ERK activation during invasion. Neoplasia (New York, N.Y.), 22(10), 470-483.

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Small Molecule Kinase Inhibitor Protocol

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Imatinib, nilotinib and dasatinib were obtained from Selleckchem (Houston, TX, USA). SNG inhibitors were obtained from Shenogen Pharma Group Ltd (Beijing, China).

Chen M., Turhan A.G., Ding H., Lin Q., Meng K, & Jiang X. (2017). Targeting BCR-ABL+ stem/progenitor cells and BCR-ABL-T315I mutant cells by effective inhibition of the BCR-ABL-Tyr177-GRB2 complex. Oncotarget, 8(27), 43662-43677.

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Cell Signaling Pathway Analysis

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RPMI-1640 and McCoy's 5A medium, fetal bovine serum (FBS), penicillin and streptomycin were purchased from Lonza Milano SRL (Milan, Italy). ZSTK474, BYL719, TGX221, AS605240, CAL101, IPI145, Imatinib, Nilotinib and GZD824 were obtained from Selleck Chemicals (Houston, TX, USA). For cell viability determination, CellTiter 96 Aqueous One Solution Cell Proliferation Assay (MTS) was purchased from Promega (Milan, Italy). Annexin V/7-ADD detection kit was from Merck-Millipore (Darmstadt, Germany). Western blot antibodies for total Akt-1, Ser473 p-Akt-1 and Thr308 p-Akt-1 were from Santa Cruz Biotechnology (Santa Cruz, CA, USA), while all the other antibodies were from Cell Signaling Technology (Danvers, MA, USA), including the rabbit secondary antibody. Chloroquine, the mouse secondary antibody and the monoclonal β-Actin antibody were purchased from Sigma Aldrich (Milan, Italy). Signals were detected using ECL Plus reagent from Perkin Elmer (Boston, MA, USA).

Ultimo S., Simioni C., Martelli A.M., Zauli G., Evangelisti C., Celeghini C., McCubrey J.A., Marisi G., Ulivi P., Capitani S, & Neri L.M. (2017). PI3K isoform inhibition associated with anti Bcr-Abl drugs shows in vitro increased anti-leukemic activity in Philadelphia chromosome-positive B-acute lymphoblastic leukemia cell lines. Oncotarget, 8(14), 23213-23227.

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FGF2 Signaling Pathway Inhibitors

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Quizartinib (AC220) was purchased from LC labs (Woburn, MA, USA). Nilotinib, PD173074 and BGJ-398 were purchased from SelleckChem (Houston, TX, USA). Imatinib was purchased from LC labs (Woburn, MA, USA). Recombinant FGF2 was purchased from Peprotech (Rocky Hill, NJ, USA).

Javidi-Sharifi N., Martinez J., English I., Joshi S.K., Scopim-Ribeiro R., Viola S.K., Edwards DK V., Agarwal A., Lopez C., Jorgens D., Tyner J.W., Druker B.J, & Traer E. (2019). FGF2-FGFR1 signaling regulates release of Leukemia-Protective exosomes from bone marrow stromal cells. eLife, 8, e40033.

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Purification and Characterization of Nilotinib

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Solvents used were puriss, with the exception of THF, which was purified by fresh distillation over Na/benzophenone. Commercially available reagents were used as received, without any further purification. Nilotinib was purchased from Selleck Chemicals, Munich, Germany. Analytical TLC was performed on commercial Merck silica gel 60 F254. Flash chromatography was carried out using silica gel 60 (230–400 mesh). HPLC experiments were performed using a Shimadzu system, consisting of a DGU-20A controller, an LC-20AD pump, an SPD-M20A photodiode-array detector and a CTO-10AS column oven. ¹H and ¹³C NMR spectra were recorded on a Brucker AMX (400/100 MHz ¹H/¹³C) spectrometer using tetramethyl silane (TMS) as an internal standard. Chemical shifts are reported in ppm (δ) referenced to TMS, coupling constants J in Hz. High-resolution ESI mass spectra were measured on a Thermo Fisher Scientific LTQ ORBITRAP/LC−MS system. Elemental analyses were performed on a Heraeus CHN-Rapid Analyser.

Pantazi D., Ntemou N., Brentas A., Alivertis D., Skobridis K, & Tselepis A.D. (2019). Molecular Requirements for the Expression of Antiplatelet Effects by Synthetic Structural Optimized Analogues of the Anticancer Drugs Imatinib and Nilotinib. Drug Design, Development and Therapy, 13, 4225-4238.

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Immunotoxins and Small Molecule Inhibitors

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Three immunotoxins, HB21PE40, HA22 and SS1P were produced recombinantly in E. coli as described previously [38 (link)]. Nilotinib and everolimus were purchased from Selleck Chemicals LLC (Houston, TX), dissolved in dimethyl sulphoxide (DMSO) at 10 mmol/L stock concentration, and stored frozen at −80°C. Salinomycin was from Sigma-Aldrich (St. Louis, MO) and was handled similarly.

Antignani A., Mathews Griner L., Guha R., Simon N., Pasetto M., Keller J., Huang M., Angelus E., Pastan I., Ferrer M., FitzGerald D.J, & Thomas C.J. (2016). Chemical Screens Identify Drugs that Enhance or Mitigate Cellular Responses to Antibody-Toxin Fusion Proteins. PLoS ONE, 11(8), e0161415.

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Compound Screening for Drug Discovery

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Chemicals used in this study were from the following sources: nilotinib (Selleckchem, S1033), imatinib (Selleckchem, S2475), torin1 (Selleckchem, S2827), Gefitinib (MedChemExpress, HY-50895), GSK621 (Sigma, SML2003), EX229 (MedChemExpress, HY-112769), BC1618 (MedChemExpress, HY-134656), bafilomycin A1 (Selleckchem, S1413), MK2206 (Selleckchem, S1078), SRT1720 (Apexbio, A4180–10), FR180204 (Selleckchem, S7524), PLX-4720 (Selleckchem, S1152). PEG300 (Selleckchem, S6704), DMSO (Sigma, D2650).

Zheng W., Chang I.C., Limberis J., Budzik J.M., Zha B.S., Howard Z., Chen L, & Ernst J.D. (2023). Mycobacterium tuberculosis resides in lysosome-poor monocyte-derived lung cells during chronic infection. bioRxiv.

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Small Molecule Compound Preparation

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All compounds (imatinib, nilotinib, dasatinib, ponatinib, crizotinib, and asciminib) used in this study were purchased from Selleck Chemicals (Houston, TX, USA) and were diluted in DMSO to a 1000× stock solution, which then were diluted to 1× working concentrations for the experiments. For the in vivo experiments, the compounds were diluted to working concentration in 0.5% methyl-cellulose (Methocel® 65HG, Fluka, Darmstadt, Germany).

Mian A.A., Haberbosch I., Khamaisie H., Agbarya A., Pietsch L., Eshel E., Najib D., Chiriches C., Ottmann O.G., Hantschel O., Biondi R.M., Ruthardt M, & Mahajna J. (2021). Crizotinib acts as ABL1 inhibitor combining ATP-binding with allosteric inhibition and is active against native BCR-ABL1 and its resistance and compound mutants BCR-ABL1T315I and BCR-ABL1T315I-E255K. Annals of Hematology, 100(8), 2023-2029.

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