The antibodies used in this study included NRF2 (16396-1-AP), Histone H3 (17168-1-AP), SLC7A11 (26864-1-AP), HMOX-1 (10701-1-AP), and GAPDH (16396-1-AP) (ProteinTech Group, IL, United States). 4-Hydroperoxy cyclophosphamide (4HC) was purchased from TCR (39800-16-3, Santa Cruz Biotechnology, TA, United States). Cyclophosphamide (CTX), Hemin, zinc protoporphyrin (ZNPP), ferrostatin-1 (Fer1), and deferoxamine mesylate salt (DFO) were purchased from Sigma–Aldrich (Sigma–Aldrich, MO, United States).
Deferoxamine mesylate salt dfo
Manufactured by Merck Group
Sourced in United States
Deferoxamine mesylate salt (DFO) is a laboratory reagent used in various research applications. It is a siderophore, a type of molecule that can chelate and bind to iron ions. DFO is commonly used in scientific research to study iron homeostasis and transport, as well as for other applications where the sequestration of iron is required.
Automatically generated - may contain errors
Lab products found in correlation
Erastin, by Merck Group (3 mentions)
Ferrostatin 1 fer 1, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Zinc protoporphyrin, by Merck Group (1 mentions)
4 hydroperoxy cyclophosphamide, by Santa Cruz Biotechnology (1 mentions)
Isoniazid, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Cloroquine cq, by Merck Group (1 mentions)
6 hydroxy 2 5 7 8 tetra methylchromane 2 carboxylic acid trolox, by Cayman Chemical (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Rapamycin, by Thermo Fisher Scientific (1 mentions)
Bafilomycin a1 baf, by Merck Group (1 mentions)
Torin 1, by Cell Signaling Technology (1 mentions)
Deferiprone dfp, by Merck Group (1 mentions)
Ferric ammonium citrate, by Merck Group (1 mentions)
Spermine, by Merck Group (1 mentions)
Cyquant direct cell proliferation assay, by Thermo Fisher Scientific (1 mentions)
Enterobactin, by Merck Group (1 mentions)
Chrysophanol, by Merck Group (1 mentions)
P jnk, by ABclonal (1 mentions)
19 protocols using deferoxamine mesylate salt dfo
1
Antioxidant Modulation in Oxidative Stress
2
TEM Analysis of M. tuberculosis Stress
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For TEM analysis of M. tuberculosis cells under different stress conditions, M. tuberculosis culture in 7H9T medium at O.D600 0.3–0.5 was treated with H2O2 (Merk) at different concentrations, ranging from 21 to 210 mM for 48 h at 37°C and selected 21 mM H2O2-treated samples for electron microscopy (Voskuil et al., 2011 (link)). For iron deficiency, M. tuberculosis isolates were cultured in the presence of deferoxamine mesylate salt (DFO) (Sigma–Aldrich) at final concentrations of 100, 250, and 500 μM in 7H9T medium until the O.D600 reached 0.3–0.5, with the 100 and 500 μM DFO-treated samples processed for electron microscopy (Pal et al., 2015 (link)). For isoniazid treatment, M. tuberculosis isolates were grown in the presence of isoniazid (Sigma–Aldrich) in 7H9T medium at a concentration of 0.015 μg/ml until the O.D600 reached 0.3–0.5. All treated and untreated control isolates, along with about 500 μl of sputum with high density of acid fast bacilli (3+) as observed by microscopy from two pulmonary tuberculosis patients, were then processed for TEM.
3
Culturing and Differentiating Murine Macrophages
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RAW264.7 cells were cultured in Dulbecco Modified Eagle Medium (Life Technologies, Grand Island, NY, USA), 5% fetal bovine serum, 100 units/mL penicillin, and 100 mg/mL streptomycin under an atmosphere of 95% air and 5% CO2 at 37 °C. Bone marrow-derived macrophages (BMDMs) from C57BL/6 mice were isolated and differentiated as described previously [49 (link)]. Briefly, bone marrow cells (3 × 107 cells) were cultured in macrophage-differentiation medium with GM-CSF at 37 °C for 7 days. The adherent macrophages were detached from culture dishes by treatment with 5% EDTA in PBS, followed by scraping with a sterile cell scraper. The resuspended cells were then directly seeded on cell culture plates for other experiments.
mitoNEET inhibitor, NL-1, was purchased from (Merck Millipore, Billerica, MA, USA, 475825). Deferoxamine mesylate salt (DFO) was purchased from (Sigma-Aldrich, St Louis, MO, D9533). Lipopolysaccharides from Escherichia coli O26:B6, γ-irradiated, BioXtra, suitable for cell culture (Sigma-Aldrich, St Louis, MO, L2654).
mitoNEET inhibitor, NL-1, was purchased from (Merck Millipore, Billerica, MA, USA, 475825). Deferoxamine mesylate salt (DFO) was purchased from (Sigma-Aldrich, St Louis, MO, D9533). Lipopolysaccharides from Escherichia coli O26:B6, γ-irradiated, BioXtra, suitable for cell culture (Sigma-Aldrich, St Louis, MO, L2654).
4
Ferroptosis Modulation in Gynecologic Oncology
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Ferlixit (62.5 mg/5 ml, sodium ferric gluconate complex in sucrose, SANOFI) and cisplatin (50 mg/100 ml, SANDOZ) were obtained from the outpatient pharmacy at the Unit of Gynecologic Oncology, Magna Graecia University of Catanzaro; deferoxamine mesylate salt (DFO), erastin, ferrostatin-1 (Fer-1), hemin and cloroquine (Cq) were purchased from Sigma Aldrich (Sigma-Aldrich, St. Louis, MO, USA); the antioxidant (±)-6-hydroxy-2,5,7,8-tetra-methylchromane-2-carboxylic acid (trolox) was ordered from Cayman Chemical (Cayman Chemical Company, Ann Arbor, USA). Cells were seeded in a 6-well plate in serum-free medium. Each compound was used at the following final concentrations: ferlixit at 100 µM and 250 µM for 1, 8, 24 or 48 h; cisplatin at 150 μM for 24 h; DFO at 200 µM for 24 h; erastin at 8 µM and 25 µM for 1, 8 or 24 h; Fer-1, hemin and Cq at 10 µM for 8, 24 or 48 h; and trolox at 200 μM for 8 h. Treatments were performed at least three times on independent biological replicates.
5
Ferroptosis Induction and Inhibition Assay
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Rapamycin was purchased from Thermo Fischer Scientific (Cat#FSBBP2963-1). Torin 1 was purchased from Cell Signalling Technologies (Cat#14379S). RSL3 was purchased from Jomar Bioscience (Cat# S8155). Erastin (Cat#E7881), ferric ammonium citrate (FAC) (Cat#F5879), deferoxamine mesylate salt (Dfo) (Cat#D9533), deferiprone (Dfp) (Cat#379409), bafilomycin A1 (Baf) (Cat# B1793) and MG-132 (ready made solution) (Cat# M7449) were purchased from Sigma Aldrich. All other reagents were purchased from Sigma Aldrich unless otherwise stated.
6
Bacterial Growth Dynamics with Iron Modulators
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Bacteria were grown on blood agar at 37°C for 24 hours, adjusted to an optical density (OD) at 650 nm of 0.01 or 0.05 in 15 ml of Lennox LB broth or iron-free M9 minimal medium, and incubated with or without the ferric ammonium citrate (FAC, 100 μM, Sigma- Aldrich, Taufkirchen, Germany), deferoxamine mesylate salt (DFO, 50 μM, Sigma-Aldrich), hepcidin-1 trifluoroacetate salt (1 μg/ml, Bachem, Bubendorf, Switzerland), or the corresponding vehicle (A. bidest) at 37°C with shaking at 140 rpm. In accordance with the time points used for the antibiotic protection and lactate dehydrogenase assay, the OD650 of bacterial cultures were measured at 0, 6, and 24 hours. Bacterial growth was determined as described [32 (link)].
7
Cell Proliferation Assays for Transduced MSCs
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For cell counting experiments, transduced MSCs were plated in triplicate for each time point and condition in 12-well plates (10,000 cells per well). The day after (day 0) and at day 3, media was changed to fresh MEM-α + 10% FBS, with or without supplements. Spermine (cat# 55513-100MG), Ammonium iron (III) citrate (FAC; cat# F5879-100G) and deferoxamine mesylate salt (DFO; cat# D9533-1G) were purchased from Sigma-Aldrich. At the indicated time points, cells to count were lifted using Trypsin and counted using Trypan Blue exclusion dye and hemocytometer.
As an orthologous method to measure cell proliferation we used a CyQUANT™ Direct Cell Proliferation Assay (Thermo Fisher, cat# C35011). Here, transduced MSCs were plated in triplicate into 96-well plates (500 cells per well). At each indicated time point, a plate was frozen at −80 °C. After the last time point, plates were treated with lysis buffer and a DNA-binding fluorescent dye as described by the manufacturer. Plates were measured by fluorescence at 520 nm emission.
As an orthologous method to measure cell proliferation we used a CyQUANT™ Direct Cell Proliferation Assay (Thermo Fisher, cat# C35011). Here, transduced MSCs were plated in triplicate into 96-well plates (500 cells per well). At each indicated time point, a plate was frozen at −80 °C. After the last time point, plates were treated with lysis buffer and a DNA-binding fluorescent dye as described by the manufacturer. Plates were measured by fluorescence at 520 nm emission.
8
Campylobacter Ferric Enterobactin Growth Assay
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As previous studies demonstrated that Campylobacter used ferric enterobactin as the sole source of iron during growth promotion assays (56 (link)), we measured the growth of C. jejuni strains as previously described (57 (link)). Briefly, a culture grown overnight on MH agar was resuspended in MH broth to an OD600 of 0.1. C. jejuni cells were grown in a disposable glass tube to log phase. Deferoxamine mesylate salt (DFO) (Sigma-Aldrich, MO, USA), a chelator, was added to melted MH agar at a final concentration of 20 μM. The cells were mixed with DFO-containing MH agar and adjusted to approximately 107 CFU/mL. Each sample mixture was poured into petri dishes for solidification. A sterile disk containing 25 μL of enterobactin (2 mM) (Sigma-Aldrich, MO, USA) was placed on the surface of the agar in each dish. Autoclaved distilled water was used instead of enterobactin as a negative control.
9
Chemical Compounds for Research
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Deferoxamine mesylate salt (DFO), Z-FF-FMK, and cryptotanshinone (CT) were purchased from Sigma-Aldrich (St Louis, MO, USA).
10
Small Molecule Modulation of Ferroptosis
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Erastin (Cat. #571203-78-6) was purchased from Cayman Chemical (Ann Arbor, MI). Ferrostatin-1 (Fer-1; Cat. #SML0583), (1S,3R)-RSL3 (Cat. #SML2234) liproxstain-1 (Lipro-1; Cat. # SML1414) and deferoxamine mesylate salt (DFO, Cat. #D9533) were from Sigma-Aldrich (St. Louis, MO). Staurosporine (Cat. #9953) was from Cell Signaling (Danvers, MA). Other chemicals used were of analytical grade.
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