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Ponatinib

Manufactured by LC Laboratories
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Ponatinib is a protein kinase inhibitor used as a laboratory research tool. It functions by inhibiting the activity of various tyrosine kinases, including BCR-ABL, FGFR, PDGFR, and SRC family kinases. Ponatinib is commonly used in cell-based and biochemical assays to study the role of these kinases in biological processes.

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Lab products found in correlation

Imatinib, by LC Laboratories (3 mentions) Pimozide, by Bio-Techne (2 mentions) Trametinib, by LC Laboratories (2 mentions) Ruxolitinib, by LC Laboratories (2 mentions) Ponatinib, by Selleck Chemicals (2 mentions) Vincristine, by Selleck Chemicals (1 mentions) Dasatinib, by Selleck Chemicals (1 mentions) Doxorubicin, by Merck Group (1 mentions) Venetoclax, by Selleck Chemicals (1 mentions) Stattic, by Selleck Chemicals (1 mentions) Obatoclax mesylate, by Selleck Chemicals (1 mentions) Pd173074, by Selleck Chemicals (1 mentions) Nintedanib, by LC Laboratories (1 mentions) Paclitaxel, by Bristol-Myers Squibb (1 mentions) Vandetanib, by Selleck Chemicals (1 mentions) Cabozantinib, by Selleck Chemicals (1 mentions) Lenvatinib, by Selleck Chemicals (1 mentions) Antibodies to phospho ret tyr905, by Cell Signaling Technology (1 mentions) Anti cytokeratin antibody, by Agilent Technologies (1 mentions) Cleaved parp, by Cell Signaling Technology (1 mentions)

9 protocols using ponatinib

1

Inhibitor-based Signaling Pathway Analysis

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Imatinib was purchased from LC Laboratories. Stock solutions were prepared in sterile water at 10 mmol/L and stored at −20°C. Small molecule inhibitor of DUSP6 (2-benzylidene-3-(cyclohexylamino)-1-lndanone hydrochloride; BCI) and the ABL1 activator DPH were purchased from Sigma-Aldrich. BCI-215 was a gift from Dr. Andreas Vogt. The MEK inhibitor (trametinib), the JAK inhibitor (ruxolitinib) and ponatinib were purchased from LC Laboratories. Pimozide (STAT5 inhibitor) was purchased from TOCRIS. Stock solutions were prepared in DMSO at 10 mmol/L and stored at −20°C. For in vivo experiments, compounds were dissolved in NMP:PEG300 (1:9). Ph-like ALL cells (Fig 2e) were treated with ruxolitinib for 20 hours and then processed for Western blotting.
Chan L.N., Murakami M.A., Robinson M.E., Caeser R., Sadras T., Lee J., Cosgun K.N., Kume K., Khairnar V., Xiao G., Ahmed M.A., Aghania E., Deb G., Hurtz C., Shojaee S., Hong C., Pölönen P., Nix M.A., Chen Z., Chen C.W., Chen J., Vogt A., Heinäniemi M., Lohi O., Wiita A.P., Izraeli S., Geng H., Weinstock D.M, & Müschen M. (2020). Signaling input from divergent pathways subverts B-cell transformation. Nature, 583(7818), 845-851.
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2

Quantification of Signaling Proteins

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Antibodies to phospho-RET(Y905) (cat. #: 3221), cleaved-PARP (cat. #: 9541), and Flag (cat. #: 2368) were purchased from Cell Signaling Technology (Danvers, MA). Selpercatinib and pralsetinib were from Chemieteck (Indianapolis, IN, USA). The purity of selpercatinib was >99.5% determined by HPLC, NMR, and quantitative elemental analysis. The purity of pralsetinib was >99% determined by achiral and chiral HPLCs, NMR, and quantitative elemental analysis. Ponatinib was from LC Laboratories (Woburn, MA). The purity was >99% determined by HPLC and quantitative elemental analysis.
Melo-Oliveira R., Oliveira I.C, & Coruzzi G.M. (1996). Arabidopsis mutant analysis and gene regulation define a nonredundant role for glutamate dehydrogenase in nitrogen assimilation.. Proceedings of the National Academy of Sciences of the United States of America, 93(10), 4718-4723.
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3

NADPH Synthesis and Purification

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All reagents, unless otherwise stated, were purchased from Sigma-Aldrich (Poole, United Kingdom). NADPH was obtained from Melford Laboratories (Ipswich, United Kingdom), ponatinib was purchased from LC Laboratories (Woburn, MA), and 2,3,7,8-tetrachlorodibenzodioxin (TCDD) was purchased from Toronto Research Chemicals (Toronto, Canada). The Nanosep centrifugal device with Omega ultrafiltration membrane molecular weight cutoff of 10 kDa was obtained from Pall Corporation (East Hills, NY).
Lin D., Kostov R., Huang J.T., Henderson C.J, & Wolf C.R. (2017). Novel Pathways of Ponatinib Disposition Catalyzed By CYP1A1 Involving Generation of Potentially Toxic Metabolites. The Journal of Pharmacology and Experimental Therapeutics, 363(1), 12-19.
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4

Pharmacological Agents Acquisition Protocol

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Ponatinib was purchased from LC Laboratories (Woburn, MA, USA). Trabectedin was obtained from PharmaMar (Colmenar Viejo, Spain). Cisplatin was kindly provided by the Institute of Inorganic Chemistry, University of Vienna. Ponatinib, dasatinib, obatoclax mesylate, venetoclax, vincristine, PD173074, and stattic were obtained from Selleckchem (EUBIO, ANDREAS KÖCK e.U., Vienna, Austria). Nintedanib and imatinib were purchased from LC Laboratories (Woburn, MA, USA). Doxorubicin was acquired from Sigma-Aldrich (St. Louis, MO, USA). Paclitaxel was procured from Bristol Myers Squibb (New York City, NY, USA).
Ghanim B., Baier D., Pirker C., Müllauer L., Sinn K., Lang G., Hoetzenecker K, & Berger W. (2022). Trabectedin Is Active against Two Novel, Patient-Derived Solitary Fibrous Pleural Tumor Cell Lines and Synergizes with Ponatinib. Cancers, 14(22), 5602.
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5

Analyzing RET and Apoptosis Signaling

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Antibodies to phospho-RET(Tyr905) and cleaved-PARP were from Cell Signaling Technology, antibodies to Flag-tag and β-actin were from Sigma, anti-RET antibody was from Santa Cruz Biotech. Anti-TTF1 (ab137061) antibody was from Abcam. Anti-cytokeratin antibody was from Dako (cat. No. Z0622). Ponatinib was from LC Laboratories. Cabozantinib, vandetanib, and lenvatinib were from Selleckchem.
Huang Q., Schneeberger V.E., Luetteke N., Jin C., Afzal R., Budzevich M.M., Makanji R.J., Martinez G.V., Shen T., Zhao L., Fung K.M., Haura E.B., Coppola D, & Wu J. (2016). Preclinical Modeling of KIF5B-RET Fusion Lung Adenocarcinoma. Molecular cancer therapeutics, 15(10), 2521-2529.
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6

Inhibitor-based Signaling Pathway Analysis

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Imatinib was purchased from LC Laboratories. Stock solutions were prepared in sterile water at 10 mmol/L and stored at −20°C. Small molecule inhibitor of DUSP6 (2-benzylidene-3-(cyclohexylamino)-1-lndanone hydrochloride; BCI) and the ABL1 activator DPH were purchased from Sigma-Aldrich. BCI-215 was a gift from Dr. Andreas Vogt. The MEK inhibitor (trametinib), the JAK inhibitor (ruxolitinib) and ponatinib were purchased from LC Laboratories. Pimozide (STAT5 inhibitor) was purchased from TOCRIS. Stock solutions were prepared in DMSO at 10 mmol/L and stored at −20°C. For in vivo experiments, compounds were dissolved in NMP:PEG300 (1:9). Ph-like ALL cells (Fig 2e) were treated with ruxolitinib for 20 hours and then processed for Western blotting.
Chan L.N., Murakami M.A., Robinson M.E., Caeser R., Sadras T., Lee J., Cosgun K.N., Kume K., Khairnar V., Xiao G., Ahmed M.A., Aghania E., Deb G., Hurtz C., Shojaee S., Hong C., Pölönen P., Nix M.A., Chen Z., Chen C.W., Chen J., Vogt A., Heinäniemi M., Lohi O., Wiita A.P., Izraeli S., Geng H., Weinstock D.M, & Müschen M. (2020). Signaling input from divergent pathways subverts B-cell transformation. Nature, 583(7818), 845-851.
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7

Evaluating Combination Treatments for CML-LICs

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To determine colony‐forming capacity after a combination treatment of EW‐7197 plus IM, or EW‐7197 plus ponatinib, we cocultured freshly isolated CML‐LICs on OP‐9 stromal cells in the presence of DMSO or EW‐7197 for 24 h.18 Cells were then treated with additional DMSO, 1 μM IM (LC Laboratories, Woburn, MA), or 1 μM ponatinib (AP24534; Selleck Chemicals) and cultured for another 2 days (total, 3 days). Colonies were counted 7 days later as previously described.18
Naka K., Ishihara K., Jomen Y., Jin C.H., Kim D., Gu Y., Jeong E., Li S., Krause D.S., Kim D., Bae E., Takihara Y., Hirao A., Oshima H., Oshima M., Ooshima A., Sheen Y.Y., Kim S, & Kim D. (2016). Novel oral transforming growth factor‐β signaling inhibitor EW‐7197 eradicates CML‐initiating cells. Cancer Science, 107(2), 140-148.
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8

Quantification of Vandetanib and Ponatinib in Rat Liver Microsomes

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All solvents were of HPLC grade and reference powders were of analytical grade. Vandetanib and ponatinib were procured from LC Laboratories (Woburn, MA, USA). Formic acid, ammonium formate, and ACN were procured from Sigma-Aldrich (West Chester, PA, USA). HPLC water grade was generated by in house Milli-Q plus purification system (Millipore, Waters, USA). Preparation of RLMs was done in house using Sprague Dawely rats [11 (link)]. Human plasma was kindly gifted by King Khalid University Hospital (Riyadh, KSA). After informed consent was gotten, fasting blood samples were taken and plasma was separated and stored at −70 °C.
Amer S.M., Kadi A.A., Darwish H.W, & Attwa M.W. (2017). Liquid chromatography tandem mass spectrometry method for the quantification of vandetanib in human plasma and rat liver microsomes matrices: metabolic stability investigation. Chemistry Central Journal, 11, 45.
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9

FDA-Approved Drug Library Screening

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The Food and Drug Administration–approved drug library was provided by the National Cancer Institute Development Therapeutics Program (https://dtp.cancer.gov/organization/dscb/obtaining/available_plates.htm). Ponatinib and rapamycin were purchased from LC Laboratories. Wortmannin was purchased from Selleckchem. An equal volume of dimethyl sulfoxide (DMSO) was used as control/vehicle.
Li X., Cai Y., Goines J., Pastura P., Brichta L., Lane A., Le Cras T.D, & Boscolo E. (2019). Ponatinib Combined With Rapamycin Causes Regression of Murine Venous Malformation. Arteriosclerosis, Thrombosis, and Vascular Biology, 39(3), 496-512.
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