RPMI 1640 medium, penicillin, streptomycin, sodium pyruvate, and fetal calf serum were purchased from Gibco (Invitrogen Life Technologies, Waltham, MA, USA). Sodium fluoride, sodium orthovanadate, aprotinin, leupeptin, phenylmethylsulfonyl fluoride, phorbol 12-myristate 13-acetate, SB202190, Triton X-100, Dasatinib, Bosutinib and alexidine were purchased from Sigma. Imatinib mesylate was purchased from Enzo Life Sciences, nilotinib from Selleck Chemicals, and ponatinib from Clinisciences. The antibodies used in these studies are listed in Table 1 .
Imatinib mesylate
Manufactured by Enzo Life Sciences
Sourced in United States
Imatinib mesylate is a protein kinase inhibitor used as a reference standard or analytical tool in research. It is a white to off-white crystalline powder.
Automatically generated - may contain errors
Lab products found in correlation
Leupeptin, by Merck Group (3 mentions)
Sodium orthovanadate, by Merck Group (3 mentions)
Sodium fluoride, by Merck Group (3 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Nilotinib, by Selleck Chemicals (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Phenylmethylsulfonyl fluoride, by Merck Group (2 mentions)
Aprotinin, by Merck Group (2 mentions)
Dasatinib, by Merck Group (1 mentions)
Bosutinib, by Merck Group (1 mentions)
Sb202190, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Agarose a, by Bio Basic (1 mentions)
Dna ladder, by Solis BioDyne (1 mentions)
Mbo 1, by Promega (1 mentions)
Ethanol, by Merck Group (1 mentions)
7 protocols using imatinib mesylate
1
Stimulation and Inhibition Pathway Analysis
2
Imatinib and Metabolites Quantification
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Imatinib mesylate (Enzo, NY, USA), N-des methyl imatinib, Pyridine-N-oxide imatinib, palonosetron hydrochloride were purchased from Santa Cruz Biotechnology (TX, USA), methanol for HPLC 99.9%, 2-propanol (Riedel-deHaën, Honeywell, Germany), formic acid for mass spectrometry, 98%, ethanol (Sigma-Aldrich, Steinheim, Germany), Agarose A (Biobasic, Ontario, Canada), Tris-acetate -EDTA(TAE) (Invitrogen Life Technologies, NY, USA). In addition, DNA ladder (Solis BioDyne, Estonia), PCR Master Mix kit (2X) (Thermo Scientific, IL, USA), Restriction enzymes: RsaI, Acl I, Dde I, Mbo I, AluI (Promega, Madison, MI, USA), and Bse3D I, Bsu R I, Bst 4C I, Bse 3D I (SibEnzyme Ltd, Russia) were obtained.
3
Comprehensive Compound Procurement for Research
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H-89 dihydrochloride, D4476, IC261, LY-364947, DAPH 2, nafoxidine hydrochloride, 1,3-di-o-tolylguanidine, naftifine hydrochloride, opipramol, rifampicin, kanamycin, and the library of pharmacologically active compounds (LOPAC) were purchased from Sigma-Aldrich, Zwijndrecht, the Netherlands. Hygromycin B was acquired from Life Technologies-Invitrogen, Bleiswijk, the Netherlands. Imatinib mesylate was from Enzo Life Sciences, Raamsdonksveer, the Netherlands. VEGFR2 kinase inhibitor I and ampicillin were purchased from Calbiochem Merck-Millipore, Darmstadt, Germany. Pazopanib HCl, AT9283, and linifanib (ABT-869) were acquired from Selleck Chemicals, Munich, Germany. Quizartinib was purchased from MedChemExpress, Stockholm, Sweden. Santa Cruz BioTechnology, Heidelberg, Germany was the supplier of PDGFR tyrosine kinase inhibitor III. Dovitinib (TKI-258, CHIR-258) was from APExBIO, Houston, TX, USA. The siKinome library was acquired from Thermo Fisher Dharmacon, Waltham, MA, USA.
4
Culturing and Treatment of K562 Cells
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CML cell line K562 was obtained from ATCC (cat # CCL-243). Cryopreserved cells were thawed and grown at 37°C and 5% CO2 in RPMI medium supplemented with 15% fetal bovine serum, and 2% L-glutamine. Cells were passaged 3 times per week to maintain exponential growth. Imatinib mesylate (Enzo Life Sciences cat # ALX-270-492) was dissolved in DMSO to 100 mM master stock, aliquoted, and stored at −20°C.
5
Evaluating Imatinib and Ambrisentan Responses
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Imatinib Mesylate (Enzo life sciences, UK; Cat. no. ALX-270-492-M025) and Ambrisentan (Astra Zeneca) were added directly to the cell culture medium at clinically relevant concentrations of 10 µM67 (link) and 1.25 nM10 , respectively. BRD4 inhibitor, AZD5153 (Astra Zeneca) was added at the concentration of 16.5 nM, based on the pre-clinical use of BET inhibitors12 (link) and the in vitro dose-response experiments. Cells were incubated with the inhibitors for 24 h under normoxic (21% O2) or hypoxic (2% O2) conditions accordingly.
6
Cell Signaling Pathway Inhibitor Assay
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Imatinib mesylate was purchased from Enzo Life Sciences. BI-2536, Rigosertib, GSK-461363 and volasetib were purchased from Selleckchem. RPMI 1640 medium, and fetal calf serum (FCS) were from life technologies. Sodium fluoride, sodium orthovanadate, phenyl-methyl-sulfonyl fluoride (PMSF), aprotinin, leupeptin were purchased Sigma- Aldrich. Anti-HSP60 and anti-HSP90 were purchased from Santa Cruz Biotechnology. HRP conjugated anti-mouse and anti-goat antibodies were from Dakopatts. Anti-PARP, anti-phospho-Plk1, anti-caspase 3, anti-caspase 9, anti-Tubulin, anti-phospho-CDC25 and peroxydase-conjugated anti-rabbit antibodies were obtained from Cell Signaling Technology. Anti-Plk1 was purchased from Abcam.
7
Cell Culture Reagents and Materials
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RPMI 1640 medium, penicillin, streptomycin, sodium pyruvate, and fetal calf serum were purchased from Gibco (Invitrogen Life Technologies, Waltham, MA, USA). Sodium fluoride, sodium orthovanadate, aprotinin, leupeptin, phenyl-methylsulfonyl fluoride, and Triton X-100 were purchased from Sigma (St. Louis, MO, USA). Imatinib mesylate was purchased from Enzo Life Sciences (Farmingdale, NY, USA); nilotinib, from Selleck Chemicals (Houston, TX, USA); and doxycycline, from Calbiochem (Darmstadt, GERMANY). Validated small-interfering RNAs (siRNAs) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). The antibodies used in these studies are listed in Table 3 .
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