AZD-1775 was purchased from MedChemexpress. Clofarabine, doxorubicin, imatinib, and ponatinib were obtained from Sigma-Aldrich. Bosutinib (Bos) was purchased from Tocris, and Bosutinib isomer (Bos-I) was purchased from LC Labs. Human B and T cell precursor ALL (B/T-ALL) cell lines (B-ALL: BV-173, SUP-B15, REH, NALM-6, NALM-19; T-ALL: MOLT-4, RPMI-8402, CCRF-CEM) were cultured in RPMI-1640 (Invitrogen) with 1% l -glutamine (Sigma-Aldrich), penicillin (100 U/ml, Gibco), and streptomycin (100 μg/ml, Gibco) supplemented with 10–20% fetal bovine serum (FBS, Gibco). All the cell lines were purchased from Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH (DSMZ) website (https://www.dsmz.de ).
Ponatinib
Manufactured by Merck Group
Sourced in United States
Ponatinib is a laboratory research product manufactured by Merck Group. It is a small molecule inhibitor that targets multiple tyrosine kinases. The core function of Ponatinib is to serve as a tool for scientific research and investigation in relevant laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Imatinib, by Merck Group (3 mentions)
Dmso control, by Merck Group (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
L glutamine, by Merck Group (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Clofarabine, by Merck Group (1 mentions)
Bv173, by Thermo Fisher Scientific (1 mentions)
Azd1775, by MedChemExpress (1 mentions)
Docetaxel, by Merck Group (1 mentions)
Mouse anti myc mab, by Santa Cruz Biotechnology (1 mentions)
Sunitinib, by Merck Group (1 mentions)
Axitinib, by Merck Group (1 mentions)
Rabbit anti gfp polyclonal ab, by Novus Biologicals (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Ethanol, by Carl Roth (1 mentions)
Annexin 5 fitc antibody, by BioLegend (1 mentions)
5 protocols using ponatinib
1
Evaluating Targeted Therapies for B/T-ALL
2
Biochemical Experiments with CLIP-170 Antibodies
3
Cell Cycle Analysis of Ponatinib Treatment
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A total of 3 × 105 cells were seeded in 6-well plates. On the next day, cells were treated with ponatinib at 10 × IC50 or DMSO control (Sigma-Aldrich, St. Louis, MO, USA) for 24, 48, and 72 h. At each time point, cells were harvested and fixed with 85% ice-cold ethanol (Carl Roth GmbH, Karlsruhe, Germany). After incubation for 30 min at 4 °C, cells were centrifuged at 300× g for 5 min at 4 °C and ethanol was carefully removed. Cells were then washed with 1 mL ice-cold DPBS twice. Thereafter, cells were resuspended in 500 μL DPBS and 5 μL RNAse A (Lucigen, Middleton, WI, USA) was added to the cells and incubated for 30 min at 4° C. Prior to measurement, 1 µL propidium iodide (PI) (Sigma-Aldrich, St. Louis, MO, USA) was added at a final concentration of 50 µg/mL. Cells were analyzed by LSR II flow cytometer (BD Biosciences, San Jose, CA, USA) and cell cycle distribution was analyzed using the software FlowJo (version 10.6.1, FlowJo LLC, Ashland, OR, USA).
4
Apoptosis Quantification by Flow Cytometry
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A total of 3 × 105 cells were seeded in 6-well plates. On the following day, cells were treated with ponatinib at 10 × IC50 or DMSO control (Sigma-Aldrich, St. Louis, MO, USA) for 24, 48, and 72 h, respectively. At each time point, cells were collected from each well and transferred into a 15 mL Falcon tube (Corning Science Mexico SA de CV, Tamaulipas, Mexico). Subsequently, cells were centrifuged at 300× g for 5 min at 4 °C. Then, the supernatant was carefully discarded and the cell pellets were resuspended and washed twice with 1 mL ice-cold DPBS. Thereafter, cells were washed with 1 mL 1 × annexin V binding buffer. After centrifugation, the supernatant was removed and the cell pellet was resuspended with 100 µL 1 × annexin V binding buffer. Finally, cells were stained with 2 µL annexin V FITC antibody (Biolegend, San Diego, CA, USA) and 50 µg/mL PI and incubated for 15 min in the dark at 4 °C. Before detection, 1 × annexin V binding buffer was added to the cells to a total volume of 500 μL. Cells were then measured by flow cytometry within 1 h and analyzed using the software FlowJo (version 10.6.1, FlowJo LLC, Ashland, OR, USA).
5
HPLC Analysis of Imatinib and Derivatives
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Imatinib, nilotinib, dasatinib, ponatinib, triethylamine, methanol, and acetonitrile were purchased from Sigma-Aldrich (Milan, Italy). A Milli DI system from Millipore (Milan, Italy) was used to create high pressure liquid chromatography (HPLC)-grade water. For the chromatographic run, a mobile phase constituted by 40% of solution A (72.5% H2O + 25% CH3OH + 2.5% C6H15N), 40% C2H3N, and 20% CH3OH was used.
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