For ROS generation, tert-butyl hydroxide solution (TBH, Sigma) was used at different doses depending on the experiment; see respective figure legends. Specific GPX4 inhibitor (B6095; APExBIO) was used. Specific cell death inhibitor and ALOX12 activity inhibitor were used in the experiments: Ferrostatin-1(ferroptosis inhibitor; Xcess Biosciences) and ML-355 (ALOX12 inhibitor; Cayman Chemical) was used at different doses depending on the experiment; see respective figure legends.
Ml355
Manufactured by Cayman Chemical
Sourced in United States
The ML355 is a laboratory centrifuge designed for high-speed separation of samples. It has a maximum speed of 17,500 rpm and can accommodate a variety of rotor types to accommodate different sample sizes and types. The ML355 is suitable for applications such as cell harvesting, DNA/RNA purification, and protein precipitation.
Automatically generated - may contain errors
Lab products found in correlation
B6095, by Apexbio (2 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Dasatinib, by LC Laboratories (1 mentions)
Imatinib, by LC Laboratories (1 mentions)
Cd34 microbead kit, by Miltenyi Biotec (1 mentions)
Zileuton, by Cayman Chemical (1 mentions)
Triacsin c, by Cayman Chemical (1 mentions)
1 aminobenzotriazole, by Cayman Chemical (1 mentions)
Het0016, by Cayman Chemical (1 mentions)
Celecoxib, by Cayman Chemical (1 mentions)
Pd146176, by Cayman Chemical (1 mentions)
Msppoh, by Cayman Chemical (1 mentions)
Lipopolysaccharides (lps), by InvivoGen (1 mentions)
2 tedc, by Bio-Techne (1 mentions)
Pd146176, by Bio-Techne (1 mentions)
Bodipy 581 591 c11, by Thermo Fisher Scientific (1 mentions)
5 protocols using ml355
1
Ferroptosis Inhibition in Cellular Assays
2
Purification and Culture of BP-CML CD34+ Cells
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BP-CML CD34+ cells (5x10^6 to 10^8 per sample) were purified from bone marrow or peripheral blood of CML patients at blast phase seen at Wuhan Forth Hospital with informed consents approved by the Tongji Medical College Centralised Institutional Review Board (No. 2,017,016). Mononuclear cells were obtained by using Ficoll separation, followed by CD34+ cell selection using CD34 MicroBead kit (Miltenyi Biotec). BP-CML CD34+ and normal bone marrow (NBM) CD34+ cells (StemCell Technologies) were cultured using the same growth factors and cytokines supplemented serum-free medium as described in our previous study.17 (link) Human CML cell lines were cultured in RPMI1640 medium (Life Technologies) supplemented with 10% fetal bovine serum (Hyclone) and 2mM glutamine (Invitrogen). Imatinib (LC laboratories) was reconstituted in PBS. Dasatinib (LC laboratories) and ML355 (Cayman chemical) were dissolved in dimethylis sulfoxidum (DMSO). Antibodies against Bcr-Abl, p-Crkl, and ALOX12 and corresponding secondary HRP-conjugated antibodies were purchased from Santa Cru and standard deviation z Biotechnology.
3
Biochemical Pathway Modulation
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ATP and H2O2 was purchased from Wako (Tokyo, Japan). LPS was purchased from Invivogen (San Diego, CA, USA). Aspirin was purchased from Tokyo Kasei (Tokyo, Japan). 1-Aminobenzotriazole, HET0016, Zileuton, PD146176, Celecoxib, MS-PPOH, ML355, and Triacsin C were purchased from Cayman (Ann Arbor, MI, USA).
4
Ferroptosis Inhibition in Cellular Assays
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For ROS generation, tert-butyl hydroxide solution (TBH, Sigma) was used at different doses depending on the experiment; see respective figure legends. Specific GPX4 inhibitor (B6095; APExBIO) was used. Specific cell death inhibitor and ALOX12 activity inhibitor were used in the experiments: Ferrostatin-1(ferroptosis inhibitor; Xcess Biosciences) and ML-355 (ALOX12 inhibitor; Cayman Chemical) was used at different doses depending on the experiment; see respective figure legends.
5
Lipoxygenase Inhibitors and Cellular Oxidation Assay
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Unless specified below, all chemical reagents were of research grade and were obtained from Merck (Kenilworth, NJ, USA). Those chemicals sourced from Thermo Fisher Scientific (Waltham, MA, USA) included BODIPY 581/591 C11 and the LIVE/DEAD viability reagent. The lipoxygenase inhibitors, PD146176, BW-B-70C and 2-TEDC were purchased from Tocris Bioscience (Avonmouth, Bristol, UK). ML355 and docosapentanoic acid were supplied by Cayman Chemical (Ann Arbor, MI, USA). Anti-ALOX15 antibodies were purchased from Abcam (Cambridge, MA, USA), while GE Healthcare (Chicago, IL, USA) supplied nitrocellulose membranes and Percoll. Tris-HCl was obtained from ICN Biochemicals (Castle Hill, NSW, Australia) and Hams-F10 was bought from MP Biomedicals (Irvine, CA, USA). The catalogue numbers of antibodies and details for the working solutions of each inhibitor and probe are provided in Supplementary Table S1 .
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