The antibodies used were as follows: rabbit antibody against human E-cadherin (24E10; Cell Signaling Technology, Danvers, MA, United States); rabbit antibody against human N-cadherin (GTX127345; GENETEX, Inc., Irvine, CA, United States); rabbit antibody against human vimentin (EPR3776; Abcam, Cambridge, United Kingdom); rabbit antibody against human β-actin (4967; Bioss Antibodies, Woburn, MA, United States); and HRP-conjugated secondary antibody against rabbit IgG (7074; Cell Signaling Technology). The inhibitors used were as follows: deferoxamine (DFO; D9533; Sigma-Aldrich, St. Louis, MO, United States), ferrostatin-1 (SML0583; Sigma-Aldrich), and MitoQ (B1309; Biovision, Milpitas, CA, United States).
Deferoxamine dfo d9533
Manufactured by Merck Group
Sourced in United States
Deferoxamine (DFO; D9533) is a chelating agent used in laboratory settings. It functions by binding and forming complexes with iron, a process known as chelation. This property allows for the effective removal and management of excess iron in various experimental and analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Mitoq, by Abcam (1 mentions)
Gtx127345, by GeneTex (1 mentions)
Epr3776, by Abcam (1 mentions)
Ferrostatin 1, by Merck Group (1 mentions)
Pgex 4t 1, by Cytiva (1 mentions)
Pfu dna polymerase, by Agilent Technologies (1 mentions)
Dpni, by New England Biolabs (1 mentions)
Actinomycin d, by Merck Group (1 mentions)
Proox p110 oxygen controller, by Biospherix (1 mentions)
Ab65080, by Abcam (1 mentions)
Ab86707, by Abcam (1 mentions)
Ab155282, by Abcam (1 mentions)
Ab175186, by Abcam (1 mentions)
4 protocols using deferoxamine dfo d9533
1
Antibodies and Inhibitors in Cell Research
2
Cloning and Mutagenesis of IRP1 in Expression Vectors
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The full-length IRP1 CDS was cloned from a human cDNA library and then inserted into the bacterial expression vector pGEX-4T-1 (Amersham, VYA0221) and the mammalian expression vector pcDNA4/TO-Myc (Invitrogen, V102020), respectively. The other constructs were maintained in the lab.
For site-directed mutagenesis, upstream and downstream primers were synthesized containing the desired mutations. Briefly, 15 ng wildtype plasmid as the template, the corresponding primers and Pfu DNA Polymerase (Agilent, 600135) were employed for site-directed mutagenesis PCR according to the manufacturer's protocol. The PCR product was processed with the endonuclease DpnI (New England Biolabs, R0176L) to degrade the template wildtype plasmid. The product was then transformed into competent E. coli cells, and positive clones were sequenced to confirm the mutagenesis.
The regents used in this study are following: deferoxamine (DFO, D9533), deferiprone (DFP, 379409), Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP, C2920), chloroquine (CQ, C6628), and actinomycin D (A1410) were purchased from Sigma-Aldrich. EBSS (SH30014.08) was purchased from HyClone.
For site-directed mutagenesis, upstream and downstream primers were synthesized containing the desired mutations. Briefly, 15 ng wildtype plasmid as the template, the corresponding primers and Pfu DNA Polymerase (Agilent, 600135) were employed for site-directed mutagenesis PCR according to the manufacturer's protocol. The PCR product was processed with the endonuclease DpnI (New England Biolabs, R0176L) to degrade the template wildtype plasmid. The product was then transformed into competent E. coli cells, and positive clones were sequenced to confirm the mutagenesis.
The regents used in this study are following: deferoxamine (DFO, D9533), deferiprone (DFP, 379409), Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP, C2920), chloroquine (CQ, C6628), and actinomycin D (A1410) were purchased from Sigma-Aldrich. EBSS (SH30014.08) was purchased from HyClone.
3
Oxygen-Induced Retinal Neovascularization and Pharmacological Modulation
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Postnatal day-7 (P7) mice with nursing dams were exposed to 75 ± 5% oxygen for 5 days with the incubator temperature maintained at 23 ± 2 °C. Oxygen level was continuously monitored with a ProOx P110 Oxygen Controller (BioSpherix, Parish, NY, USA). The mice were returned to room air for 5 days. To investigate the effect of chemicals on neovascularization, half of the pups were administrated with daily intraperitoneal (IP) injections of the following chemicals from P12 to P16 in separate studies; Deferoxamine (DFO, D-9533; Sigma, 50 mg/kg), Deferiprone (DFP, 379409; Sigma, 50 mg/kg), Ferrostatin-1 (Fer-1, H3149; Sigma, 5 mg/kg), MPP (Sigma, 1 mg/kg), and Ferric ammonium citrate (FAC, F5879; Sigma, 20 mg/kg) diluted in saline (50 µL). The remaining half of the pups were injected with appropriate vehicles. At P17, pups were sacrificed by CO2 inhalation for retinal wholemount preparations as described below. The chemical structures of DFO and DFP were drawn using ChemSketch software (ACD/Labs, Toronto, ON, Canada).
4
Ferroptosis Pathway Modulation Assay
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Dihydroartemisinin (DHA, D140839, purity: ≥98%), hemin (H140872) and chloroquine (CQ, C193834) were purchased from Aladdin (China). Dimethyl sulfoxide (DMSO, A503039) and ferric ammonium citrate (FAC, A500061) were got from Sangon Biotech (China). Deferoxamine (DFO, D9533) and protoporphyrin IX zinc(II) (ZnPPIX, 282820) were purchased from Sigma-Aldrich (United States). Z-VAD-FMK (HY-16658B), ferrostatin-1 (Fer-1, HY-100579) and doxorubicin hydrochloride (DOX, HY-15142) were purchased from MedChemExpress (United States). And the antibody to ACSL4 (ab155282), GPX4 (ab125066), xCT (ab175186), HO-1 (ab82585), TfR1 (ab84036), FTH1 (ab65080) and NCOA4 (ab86707) were all purchased from Abcam (United Kingdom). Anti-β-actin and goat anti-rabbit IgG H&L were purchased from Bioker (China).
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