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Ferrostatin-1 (Fer-1), a potent and selective ferroptosis inhibitor, suppresses Erastin-induced ferroptosis in HT-1080 cells (EC50=60 nM). Ferrostatin-1, a synthetic antioxidant, acts via a reductive mechanism to prevent damage to membrane lipids and thereby inhibits cell death. Ferrostatin-1 exhibits antifungal activity.
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U2OS cells are incubated with DMSO or OSMI-1 for 12 h and then pre-treated with Ferrostatin-1 (Fer-1; 2 μM) for 2 h before the treatment of RSL3 for 4 h. Cells are stained with PI and cell death is then assessed by flow cytometry.
Primary HSCs and LX-2 cells are treated with EA (LX-2 cells: 40 μM; primary HSCs: 45 μM), Ferrostatin-1 (1 μM) and ZVAD-FMK (10 μM), and cells are stained with PI (red fluorescence) to examine the dead cells.
Mitochondrial membrane potential (ΔΨm) measured by fluorescence microscopy of MCC treated with TBHP and rescued via Ferrostatin-1 (Fer-1; 1 μM) using JC-1 Mitochondrial Membrane Potential Assay. In Fer-1 and DFO group, red fluorescence (poly JC-1) is increased and green fluorescence (mono JC-1) is decreased.
After the gallic acid treatment, Fe2+, ROS, and MDA are significantly increased compared to the control group, and at the same time, the content of GSH is significantly reduced. The intervention of Ferrostatin-1 (2 μM) while treating the cells with gallic acid reversed the changes in these indicators to varying degrees.
Gallic acid affects the expression of ferroptosis-related proteins. The relative expression levels of GPX4, SLC7A11, and SIGMAR1 decreased, while the relative expression levels of ATF4 and TFR1 increased. Ferrostatin-1 (2 μM) as a ferroptosis inhibitor reverses the expression trend of these proteins.
p53 expression of HepG2 cells after treated with Fer-1 at a concentration of 0-40 nM. The addition of Fer-1 could repress the expression of p53 protein by inhibiting the ferroptosis of SFB formulation.
Acot1 protein levels in different concentrations of DOX-treated HL-1 cell with or without Fer-1 (10 μM) for 24 h. Acot1 expression is decreased at a higher concentration of DOX, while Fer-1 co-treatment partially suppressed this trend.
Ferrostatin-1 (Fer-1), a potent and selective ferroptosis inhibitor, suppresses Erastin-induced ferroptosis in HT-1080 cells (EC50=60 nM). Ferrostatin-1, a synthetic antioxidant, acts via a reductive mechanism to prevent damage to membrane lipids and thereby inhibits cell death. Ferrostatin-1 exhibits antifungal activity[1][2][3].
Inhibition of erastin-induced cell death in FRDA patient-derived fibroblast assessed as depletion of cellular ATP incubated for 12 hrs followed by erastin stimulation and measured after by luciferase-linked ATPase enzymatic assay
Inhibition of erastin-induced cell death in FRDA patient-derived fibroblast assessed as depletion of cellular ATP incubated for 12 hrs followed by erastin stimulation and measured after by luciferase-linked ATPase enzymatic assay
Anti-ferroptotic activity against Erastin-induced ferroptosis in human HT-1080 cells assessed as cell activity measured after 24 hrs by Cell Titer-Glo luminescent assay
Anti-ferroptotic activity against Erastin-induced ferroptosis in human HT-1080 cells assessed as cell activity measured after 24 hrs by Cell Titer-Glo luminescent assay
Antiferroptotic activity in human IMR32 neuroblastoma cells assessed as inhibition of erastin-induced cell death preincubated for 1 hr before erastin stimulation and measured after 13 hrs by fluorescence plate reader method
Antiferroptotic activity in human IMR32 neuroblastoma cells assessed as inhibition of erastin-induced cell death preincubated for 1 hr before erastin stimulation and measured after 13 hrs by fluorescence plate reader method
Inhibition of erastin-induced ferroptosis in human IMR32 cells preincubated for 1 hr followed by erastin stimulation and measured after 13 hrs by sytox green-based fluorescence assay
Inhibition of erastin-induced ferroptosis in human IMR32 cells preincubated for 1 hr followed by erastin stimulation and measured after 13 hrs by sytox green-based fluorescence assay
Anti-ferroptotic activity in FRDA patient-derived Lymphocyte assessed as reduction in RSL3-induced lipid peroxidation incubated for overnight followed by RSL3 stimulation and measured after 90 mins by FACS analysis
Anti-ferroptotic activity in FRDA patient-derived Lymphocyte assessed as reduction in RSL3-induced lipid peroxidation incubated for overnight followed by RSL3 stimulation and measured after 90 mins by FACS analysis
Anti-ferroptotic activity against RSL-3 induced ferroptosis in mouse RAW264.7 cells assessed as cell activity measured after 24 hrs by Cell Titer-Glo luminescent assay
Anti-ferroptotic activity against RSL-3 induced ferroptosis in mouse RAW264.7 cells assessed as cell activity measured after 24 hrs by Cell Titer-Glo luminescent assay
Ferrostatin-1 prevents erastin-induced accumulation of cytosolic and lipid ROS. Ferrostatin-1 prevents glutamate-induced neurotoxicity in organotypic rat brain slices[1].
Ferrostatin-1 (2 μM; 24 h) prevents Glutamate (5 mM)-induced neurotoxicity in a rat organotypic hippocampal slice culture (OHSC)[2].
Ferrostatin-1 inhibits lipid peroxidation, but not mitochondrial reactive oxygen species formation or lysosomal membrane permeability[2].
Ferrostatin-1 inhibits cell death in cellular models of Huntington's disease (HD), periventricular leukomalacia (PVL), and kidney dysfunction[2].
Ferrostatin-1 (1 μM; 6 h) inhibits the oxidative destruction of unsaturated fatty acids in HT-1080 cells, thus increases the number of healthy medium spiny neurons (MSNs)[3].
Pretreatment with 2 μM for 14 h followed by co-treatment with the inducer for 24 h
RSL3 (1 μM; 6 or 24 h)
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Ferrostatin-1 Related Antibodies
In Vivo
Ferrostatin-1 (5 mg/kg; ip; single dose, 30 min before glycerol injection) improves renal function in mice with rhabdomyolysis, whereas no beneficial effects were observed with the pan-caspase inhibitor zVAD or in RIPK3-deficient mice[1].
Ferrostatin-1 (0.8 mg/kg; tail vein injection) effectively alleviates LPS-induced induced acute lung injury (ALI)[4].
Ferrostatin-1 (i.p.; 5 mg/kg; C57BL/6J mice) improves renal function in mice with rhabdomyolysis[5].
Ferrostatin-1 (10 mg/kg/d, i.p., 3 d) attenuates hypoxic-ischemic brain damage-, oxygen-glucose deprivation-, or Erastin (HY-15763)-induced ferroptosis in brain of neonatal rats[6].
Ferrostatin-1 (0.655 mg/kg, i.p., 3 times a week for 6 week) exerts anti-ferroptosis effects by increasing GPX4 activity and by inhibiting lipid peroxidation in the salivary gland of ovariectomized (postmenopausal model) rats[7].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
DMSO : 125 mg/mL (476.46 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Preparing Stock Solutions
ConcentrationSolventMass
1 mg
5 mg
10 mg
1 mM
3.8117 mL
19.0585 mL
38.1170 mL
5 mM
0.7623 mL
3.8117 mL
7.6234 mL
10 mM
0.3812 mL
1.9059 mL
3.8117 mL
View the Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day. The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 900 μLCorn oil, and mix evenly.
Solubility: 0.2 mg/mL (0.76 mM); Clear solution; Need ultrasonic
For the following dissolution methods, please prepare the working solution directly.
It is recommended to prepare fresh solutions and use them promptly within a short period of time. The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution.
If precipitation or phase separation occurs during preparation,
heat and/or sonication can be used to aid dissolution.
Protocol 1
Add each solvent one by one: 50% PEG300 50% Saline
Solubility: 5 mg/mL (19.06 mM); Suspended solution; Need ultrasonic
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
Dosage
mg/kg
Animal weight (per animal)
g
Dosing volume (per animal)
μL
Number of animals
Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO+
%
+
%
Tween-80
+
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO,
. All of co-solvents are available by MedChemExpress (MCE).
, Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration:
mg/mL
Method for preparing stock solution:
mg
drug dissolved in
μL
DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take
μL DMSO stock solution, add
μL .
μL , mix evenly, next add
μL Tween 80, mix evenly, then add
μL Saline.
Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution
If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.
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