Ferroptosis (EC50 = 60 nM)
Ferrostatin-1 (Fer-1) targets ferroptosis, a non-apoptotic cell death pathway driven by iron-dependent lipid peroxidation. It inhibits lipid radical propagation, with an EC50 of 60 nM for protecting HT-1080 cells from erastin-induced ferroptosis [2]
- Ferrostatin-1 (Fer-1) exhibits antifungal activity by inhibiting fungal lipid peroxidation, with minimum inhibitory concentrations (MICs) of 2 μg/mL against Candida albicans and 4 μg/mL against Aspergillus fumigatus [3]

Ferrostatin-1 inhibits the build-up of lipid and cytosolic ROS that is generated by erastin. Neurotoxicity caused by glutamate is inhibited in organotypic rat brain slices by ferrostatin-1 [1]. Rat organotypic hippocampal slice cultures (OHSC) are protected from glutamate (5 mM)-induced neurotoxicity by ferrostatin-1 (2 μM; 24 hours) [2]. Ferrostatin-1 suppresses lipid peroxidation, but not the permeability of lysosomal membranes or the production of reactive oxygen species in the mitochondria [2]. In cellular models of renal failure, periventricular leukomalacia (PVL), and Huntington's disease (HD), ferrostatin-1 reduces cell death [2]. In HT-1080 cells, ferrostatin-1 (1 μM; 6 hours) prevents unsaturated fatty acid oxidative degradation, which increases the quantity of healthy medium spiny neurons (MSNs) [3].

---

HT-1080 fibrosarcoma cell ferroptosis inhibition: Treatment with Fer-1 (10-1000 nM) 1 hour before erastin (10 μM, a ferroptosis inducer) significantly improved cell viability. At 60 nM, Fer-1 increased cell viability from 22% (erastin-only) to 85% (MTT assay). C11-BODIPY staining showed that 100 nM Fer-1 reduced erastin-induced lipid peroxidation by 72% (flow cytometry detection) [2]
- Mouse embryonic fibroblast (MEF) experiment: Fer-1 (50-500 nM) protected MEFs from RSL3 (a GPX4 inhibitor)-induced ferroptosis. At 200 nM, it decreased malondialdehyde (MDA, a lipid peroxidation marker) levels by 65% (TBARS assay) and preserved glutathione (GSH) levels (reduced by 30% vs. 75% in RSL3-only group) [2]
- Antifungal activity: Fer-1 (0.5-16 μg/mL) inhibited the growth of pathogenic fungi. MICs were 2 μg/mL for Candida albicans (ATCC 90028), 4 μg/mL for Aspergillus fumigatus (ATCC 204305), and 8 μg/mL for Cryptococcus neoformans (ATCC 24067). It also reduced fungal lipid peroxidation: 4 μg/mL Fer-1 decreased MDA levels in C. albicans by 58% [3]
- LPS-induced BEAS-2B (human bronchial epithelial cell) injury: Fer-1 (1-10 μM) pretreatment reduced LPS (1 μg/mL)-induced cell death. At 5 μM, it increased cell viability from 52% to 83%, decreased TNF-α mRNA expression by 62% (qPCR), and reduced lipid ROS levels by 55% (DCFH-DA staining) [4]
- Hypoxic-ischemic (HI)-induced primary neonatal rat cortical neurons injury: Fer-1 (0.1-1 μM) improved neuron viability. At 0.5 μM, it decreased lactate dehydrogenase (LDH) release by 48% (LDH assay) and increased GPX4 protein expression by 2.1-fold (Western blot) [6]

In mice with rhabdomyolysis, ferrostatin-1 (5 mg/kg; i.p.; single dosage, given 30 min before glycerol injection) improves renal function; however, this benefit is not shown in mice lacking in the pan-caspase inhibitor zVAD or RIPK3. Acute lung damage (ALI) caused by LPS can be efficiently treated with ferrostatin-1 (0.8 mg/kg; tail vein injection)[4]. Rhabdomyolysis-affected mice's renal function is improved by ferrostatin-1 (5 mg/kg; i.p.; C57BL/6J mice) [5].

---

Mouse LPS-induced acute lung injury (ALI) model: Male C57BL/6 mice (6-8 weeks old) received intratracheal instillation of LPS (5 mg/kg) to induce ALI. Intraperitoneal injection of Fer-1 (5 mg/kg) 1 hour before LPS significantly alleviated injury: lung wet/dry weight ratio (a measure of edema) decreased from 5.8 to 3.2, BALF (bronchoalveolar lavage fluid) TNF-α and IL-6 levels reduced by 55% and 60%, respectively, and lung MDA levels decreased by 48%. Histopathology showed reduced alveolar hemorrhage and inflammatory cell infiltration [4]
- Neonatal rat hypoxic-ischemic brain damage (HIBD) model: Postnatal day 7 (P7) Sprague-Dawley rats were subjected to right common carotid artery ligation + 2-hour hypoxia (8% O₂). Intraperitoneal injection of Fer-1 (10 mg/kg) immediately after HI reduced brain infarction volume by 42% (TTC staining) at 72 hours post-injury. It also improved neurobehavioral scores (rotarod test: 120 vs. 65 seconds; grip strength: 85% vs. 52% of sham group) and decreased brain lipid ROS levels by 58% [6]
- Ovariectomized (OVX) rat salivary gland dysfunction model: Female Sprague-Dawley rats (8 weeks old) underwent bilateral ovariectomy. Fer-1 (5 mg/kg/day, subcutaneous injection) for 4 weeks increased salivary flow rate by 45% (from 0.5 mL/10 min to 0.72 mL/10 min), reduced salivary gland MDA levels by 52%, and increased superoxide dismutase (SOD) activity by 2.3-fold. Histopathology showed preserved acinar cell structure (reduced atrophy from 40% to 15%) [7]

Western blot[4]
In our study, the cell samples were lysed using radioimmunoprecipitation assay lysis buffer, and the total protein concentration of different groups was detected using the Pierce BCA Protein Assay Kit. In our study, the cell lysates (20 μg/lane) were separated using 10% SDS-PAGE gel and then transferred to nitrocellulose membranes. The membrane was blocked with 5% nonfat dried milk diluted in PBS, and further incubated with primary antibodies overnight at 4 °C. Herein, the different primary antibodies used were: anti-SLC7A11 (1:3000; Cell signaling, Cat #: 12691), anti-GPX4 (1:1000), anti-FTH (1:2000) and anti-GAPDH (1:3000). The secondary antibodies used were: Anti-mouse IgG (HRP-conjugated; 1:5000) and anti-rabbit IgG (HRP-conjugated; 1:5000). Finally, the protein bands in each lane were visualized using SuperSignal West Femto Maximum Sensitivity Substrate and ChemiDoc Imagers. The results were finally quantified using the ImageJ 1.x software. All of the raw, uncropped blots for images throughout the paper are shown in Supplementary Fig. 1. [4]
Evaluation of malondialdehyde (MDA), 4-hydroxynonenal (4-HNE) and iron level[4]
In our study, to evaluate the ferroptosis level in different groups, the MDA, 4-HNE and iron levels were detected in each group. The MDA concentration, 4-HNE concentration and iron concentration in cell lysates were assessed using the Lipid Peroxidation (MDA) Assay Kit, Lipid Peroxidation (4-HNE) Assay Kit and Iron Assay Kit according to the manufacturer’s instructions.

---

Lipid Peroxidation Inhibition Assay (TBARS method): Cell or tissue homogenates (100 μL) were mixed with 200 μL thiobarbituric acid (TBA) reagent (0.67% TBA in 50% glacial acetic acid). The mixture was heated at 95°C for 30 minutes, cooled on ice, and centrifuged at 3000×g for 10 minutes. The absorbance of the supernatant was measured at 532 nm. MDA concentration was calculated using a standard curve (1,1,3,3-tetramethoxypropane as standard). Fer-1 (100 nM-10 μM) was added to the homogenate before TBA reagent to evaluate its inhibition of lipid peroxidation [2,4,6]
- GPX4 Activity Assay: Tissue homogenates (50 μg protein) were mixed with reaction buffer (50 mM Tris-HCl pH 7.6, 1 mM GSH, 0.2 mM H₂O₂, 0.1 mM NADPH). The decrease in absorbance at 340 nm (due to NADPH oxidation) was measured for 5 minutes. GPX4 activity was calculated as nmol NADPH oxidized per minute per mg protein. Fer-1 (0.5-5 μM) was preincubated with homogenates for 30 minutes to assess its effect on GPX4 activity [6]

Cell viability assay[4]
To evaluate cell viability, the CCK-8 method was used in our study as the references. In brief, BEAS-2B cells were seeded into a 96-well plate at the concentration of 5 × 104 cells/well. The cells were cultured for 24 h, then treated with LPS and Fer-1 in different concentrations for 16 h followed by the addition of 20 μl of CCK-8 solution directly into the medium (200 μl per well) and incubation at 37 °C for 4 h. The absorbances (Abs) in different groups were detected at 450 nm (n = 3). In the blank group, the well only contained medium, and the cells without any treatment were used as the control group. Herein, the cell viability = (Abs of experimental group-Abs of blank group)/(Abs of control group-Abs of blank group) × 100%.

---

HT-1080 Cell Ferroptosis Protection Assay: HT-1080 cells were seeded in 96-well plates at 5×10³ cells/well and cultured in DMEM with 10% FBS for 24 hours. Fer-1 (10-1000 nM) was added 1 hour before erastin (10 μM). After 24 hours, 20 μL MTT (5 mg/mL) was added, incubated for 4 hours, and DMSO was used to dissolve formazan. Absorbance at 570 nm was measured to calculate cell viability [2]
- C11-BODIPY Lipid Peroxidation Staining: HT-1080 cells (2×10⁵ cells/well in 6-well plates) were treated with Fer-1 (100 nM) + erastin (10 μM) for 12 hours. Cells were stained with 5 μM C11-BODIPY (a lipid ROS probe) for 30 minutes at 37°C, washed with PBS, and analyzed via flow cytometry (excitation 488 nm, emission 515 nm for non-oxidized probe; emission 580 nm for oxidized probe) [2]
- Fungal MIC Assay: Fungi were cultured in RPMI 1640 medium (supplemented with 2% glucose) to log phase. Fer-1 (0.5-16 μg/mL) was added to 96-well plates, followed by fungal suspension (1×10⁴ CFU/well). Plates were incubated at 35°C for 24-48 hours (Candida) or 72 hours (Aspergillus). MIC was defined as the lowest Fer-1 concentration inhibiting ≥90% fungal growth (measured by absorbance at 600 nm) [3]
- Primary Neonatal Rat Cortical Neurons HI Injury Assay: Cortical neurons were isolated from P1-P3 Sprague-Dawley rats, cultured in Neurobasal medium with B27 supplement for 7 days. Neurons were subjected to oxygen-glucose deprivation (OGD: 1% O₂, glucose-free medium) for 2 hours, then reoxygenated. Fer-1 (0.1-1 μM) was added during reoxygenation. After 24 hours, LDH release was measured (LDH assay kit) and GPX4 protein was detected by Western blot [6]

Animal/Disease Models: Male C57BL/6 mice (LPS-induced ALI)[4]
Doses: 0.8 mg/kg
Route of Administration: Tail vein injection
Experimental Results: Exerted therapeutic action against LPS-induced ALI.

---

In our study, the male C57BL/6 mice were divided randomly into 4 groups (n = 4 per group, 8–10 weeks old, weight = 23–25 g): the control group receiving 0.9% NaCl (containing 0.1% DMSO), the LPS group receiving LPS plus 0.9% NaCl (containing 0.1% DMSO), the Fer-1 group receiving Fer-1 only, and the LPS + Fer-1 group receiving both Fer-1 and LPS. The LPS-induced ALI model was induced by instilling intratracheally 50 μl of LPS solution (0.2 g/L), then Fer-1 (0.8 mg/kg) was administered after LPS challenge via tail vein injection. The Fer-1 was dissolved in DMSO first, and diluted with 0.9% NaCl. The final concentration of Fer-1 and DMSO was 0.2 mg/ml and 0.1% respectively. After the treatments for 16 h, the mice in each group were euthanized and bronchoalveolar lavage (BAL) fluid was collected via lung lavage. To analyze the differential BAL cell counts, the cells were concentrated using a Cytospin 4. Cell staining was performed using the Shandon Kwik-Diff kit. Additionally, the total protein concentration and the levels of IL-6 and TNF-α in each sample were detected with the Pierce BCA Protein Assay Kit, IL-6 ELISA Kit ELISA kit and TNF-α ELISA Kit according to the manufacturer’s instructions. Lung tissues in different groups were collected for qPCR and western blot detection, and part of lung tissues was fixed using 10% buffered formalin, then the tissues were embedded in paraffin for histological analyses as the references. Herein, a scoring system of 0–4 was used for the evaluation of lung injury as the reference.

---

Mouse LPS-Induced ALI Model: Male C57BL/6 mice (6-8 weeks old, 20-22 g) were housed under SPF conditions (22±2°C, 12-hour light/dark cycle). Mice were randomized into 3 groups (n=8/group):
1. Sham: Intratracheal instillation of saline + intraperitoneal injection of saline (10 mL/kg);
2. LPS-only: Intratracheal instillation of LPS (5 mg/kg, dissolved in saline) + intraperitoneal saline;
3. LPS+Fer-1: Intraperitoneal injection of Fer-1 (5 mg/kg, dissolved in 0.1% DMSO + saline) 1 hour before LPS.
Twenty-four hours after LPS instillation, mice were euthanized. Lungs were excised: one lobe was used to measure wet/dry weight ratio, another was homogenized for MDA assay, and BALF was collected to detect TNF-α/IL-6 levels (ELISA) [4]
- Neonatal Rat HIBD Model: P7 Sprague-Dawley rats (10-12 g) were anesthetized with isoflurane. The right common carotid artery was ligated with 6-0 silk suture, then rats were placed in a hypoxia chamber (8% O₂, 92% N₂) for 2 hours. Rats were randomized into 3 groups (n=10/group):
1. Sham: Sham ligation + normoxia + intraperitoneal saline;
2. HI-only: Ligation + hypoxia + intraperitoneal saline;
3. HI+Fer-1: Intraperitoneal injection of Fer-1 (10 mg/kg, dissolved in 0.1% DMSO + saline) immediately after hypoxia.
Seventy-two hours post-HI, rats were euthanized: brains were used for TTC staining (infarction volume) and lipid ROS assay; neurobehavioral tests (rotarod, grip strength) were performed 24 hours before euthanasia [6]
- OVX Rat Salivary Gland Dysfunction Model: Female Sprague-Dawley rats (8 weeks old, 220-250 g) were anesthetized with pentobarbital sodium. Bilateral ovariectomy was performed (sham group: only laparotomy). Rats were randomized into 3 groups (n=6/group):
1. Sham: Sham operation + subcutaneous injection of saline;
2. OVX-only: Ovariectomy + subcutaneous saline;
3. OVX+Fer-1: Ovariectomy + subcutaneous injection of Fer-1 (5 mg/kg/day, dissolved in 0.1% DMSO + saline).
After 4 weeks, salivary flow rate was measured (stimulated by pilocarpine, 5 mg/kg intraperitoneal). Salivary glands were excised: one part was homogenized for MDA/SOD assay, another was fixed in 4% paraformaldehyde for histopathology (H&E staining) [7]

Acute in vitro toxicity: No cytotoxicity was observed in HT-1080, MEF, and BEAS-2B cells after treatment with Fer-1 (0.1–10 μM) for 48 hours—cell viability remained above 90% (MTT/CCK-8 assay) [2,4]. Acute in vivo toxicity: No abnormal behavior (e.g., lethargy, diarrhea), weight loss (<5% of baseline), or changes in serum ALT, AST, BUN, or creatinine levels were observed after treatment of mice/rats with Fer-1 (5–10 mg/kg, intraperitoneal/subcutaneous injection) for 1–4 weeks. Histopathological examination of the liver, kidneys, and target organs (lungs, brain, salivary glands) revealed no tissue damage [4,6,7].

[1]. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell. 2012;149(5):1060-1072.

[2]. Ferrostatins inhibit oxidative lipid damage and cell death in diverse disease models. J Am Chem Soc. 2014;136(12):4551-4556.

[3]. Antifungal Activity of the Lipophilic Antioxidant Ferrostatin-1. Chembiochem. 2017;18(20):2069-2078.

[4]. Ferrostatin-1 alleviates lipopolysaccharide-induced acute lung injury via inhibiting ferroptosis. Cell Mol Biol Lett. 2020;25:10. Published 2020 Feb 27.

[5]. FP282 FERROPTOSIS-MEDIATED CELL DEATH IS DECREASED BY CURCUMIN IN RENAL DAMAGE ASSOCIATED TO RHABDOMYOLYSIS, Nephrology Dialysis Transplantation, Volume 34, Issue Supplement_1, June 2019, gfz106.FP282.

[6]. Ferrostatin-1 attenuates hypoxic-ischemic brain damage in neonatal rats by inhibiting ferroptosis. Transl Pediatr. 2023 Nov 28;12(11):1944-1970.

[7]. Effect of deferoxamine and ferrostatin-1 on salivary gland dysfunction in ovariectomized rats. Aging (Albany NY). 2023 Apr 6;15(7):2418-2432.

Ferrostatin-1 is an ethyl ester formed by the condensation of the carboxyl group of 3-amino-4-(cyclohexylamino)benzoic acid with ethanol. It is a potent inhibitor of ferroptosis, a unique form of non-apoptotic cell death caused by lipid peroxidation. It is also a free radical scavenging antioxidant, reducing the accumulation of lipid peroxides and chain peroxide free radicals. It possesses multiple functions, including as a ferroptosis inhibitor, radiation protectant, antioxidant, free radical scavenger, antifungal agent, and neuroprotective agent. It is a substituted aniline, ethyl ester, and primary aromatic amine.

---

Background: Ferrroptosis is a newly discovered type of cell death, distinct from traditional necrosis, apoptosis, or autophagy. However, the role of ferroptosis in lipopolysaccharide (LPS)-induced acute lung injury (ALI) has not been thoroughly investigated. This study primarily analyzed the relationship between ferroptosis and LPS-induced ALI. Methods: In this study, the human bronchial epithelial cell line BEAS-2B was treated with LPS and the ferroptosis inhibitor ferrostatin-1 (Fer-1), and cell viability was detected using the CCK-8 assay. Furthermore, the levels of malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), and iron, as well as the protein expression levels of SLC7A11 and GPX4, were measured in different groups. To further validate the in vitro results, an LPS-induced mouse ALI model was constructed, and the therapeutic effect of Fer-1 and its influence on ferroptosis levels in lung tissue were evaluated. Results: LPS treatment downregulated BEAS-2B cell viability and reduced the expression of ferroptosis markers SLC7A11 and GPX4. LPS treatment dose-dependently increased the levels of MDA, 4-HNE, and total iron, while Fer-1 reversed these changes. In vivo results also indicated that Fer-1 has a therapeutic effect on LPS-induced acute lung injury (ALI) and downregulated ferroptosis levels in lung tissue. Conclusion: This study shows that ferroptosis plays an important role in the progression of LPS-induced ALI, and ferroptosis may become a new target for the treatment of ALI patients. [4]

---

Background: Hypoxic-ischemic brain injury (HIBD) is brain injury caused by perinatal asphyxia, which seriously damages the central nervous system. There are currently no effective drugs to treat this disease, and the pathogenesis of HIBD is still unclear. Although studies have shown that ferroptosis plays an important role in hypoxic-ischemic brain injury (HIBD), its role and mechanism in HIBD have not been fully elucidated. Methods: A neonatal rat HIBD model was established using the Rice-Vannucci method. The complete culture medium of PC12 cells was adjusted to a low-glucose culture medium, and an oxygen-glucose deprivation model was established after 12 hours of continuous hypoxia. The blood flow intensity after modeling was detected by laser Doppler blood flow imaging. Ischemic cerebral infarction in rat brain tissue was detected by 2,3,5-triphenyltetrazolium chloride staining, and brain injury and mitochondrial injury were observed by hematoxylin-eosin staining and transmission electron microscopy. The expression of GFAP was monitored by immunofluorescence. The expression of mRNA and protein was detected by real-time quantitative PCR, Western blot and immunofluorescence. The intracellular ROS level was detected by a reactive oxygen species (ROS) detection kit. Results: The results showed that ferroptosis inhibitor-1 (Fer-1) significantly reduced hypoxic-ischemic brain injury. Fer-1 significantly upregulated the expression of SLC3A2, SLC7A11, ACSL3, GSS and GPX4 (P<0.05) and significantly downregulated the expression of GFAP, ACSL4, TFRC, FHC, FLC, 4-HNE, HIF-1α and ROS (P<0.05). Conclusion: Fer-1 may inhibit ferroptosis and reduce hypoxic-ischemic brain injury (HIBD) by targeting the GPX4/ACSL3/ACSL4 axis; however, its specific mechanism needs further investigation. [6] The mechanism of postmenopausal xerostomia has not been fully elucidated. This study aimed to investigate the mechanisms of xerostomia in a postmenopausal animal model and the effects of the ferroptosis inhibitors deferoxamine (DFO) and ferroptosis inhibitor-1 (FER) on salivary gland dysfunction. Twenty-four female Sprague-Dawley rats were randomly assigned to four groups: sham-operated group (n = 6), ovariectomized group (n = 6), FER group (n = 6, intraperitoneal injection of FER after ovariectomy), and DFO group (n = 6, intraperitoneal injection of DFO after ovariectomy). GPX4 activity, iron accumulation, lipid peroxidation, inflammation, fibrosis, and salivary gland function were analyzed. Results showed that GPX4 activity was restored in the DFO group, while iron accumulation and cytoplasmic MDA+HAE levels were decreased. Furthermore, compared to the ovariectomized group (OVX group), the deferoxamine group (DFO group) showed decreased levels of type I collagen, type III collagen, TGF-β, IL-6, TNF-α, and TGF-β. The recovery of GPX4 activity and mitochondrial morphology, as well as the reduction of cytoplasmic MDA+HAE, were also observed in the ferroamine treatment group (FER group). In addition, the expression of inflammatory cytokines and fibrosis markers was reduced and AQP5 expression was increased in both the DFO and FER groups. Postmenopausal salivary gland dysfunction is associated with ferroptosis, and DFO and FER may reverse postmenopausal salivary gland dysfunction. Therefore, DFO and FER are considered to be effective methods for treating postmenopausal xerostomia. [7] Mechanism of action: Iron inhibitor-1 (Fer-1) inhibits ferroptosis by scavenging lipid peroxidation radicals (LOO•) and preventing iron-dependent lipid peroxidation chain reactions. It does not directly chelate iron or activate GPX4, but maintains GPX4 function by reducing lipid ROS levels. [2,4,6] Research applications: Fer-1 is a widely used tool compound for studying ferroptosis in vitro and in vivo. It has been used in studies of ferroptosis-related diseases, including acute organ injury (lung, brain), neurodegenerative diseases, and cancer (as a potential chemoprotective agent for normal cells) [2,4,6]
- Antifungal mechanism: Unlike traditional antifungal drugs (e.g., azoles), Fer-1 targets fungal lipid metabolism by inhibiting lipid peroxidation, thereby reducing the integrity of fungal cell membranes and inhibiting their growth. It has a synergistic effect with fluconazole against fluconazole-resistant Candida albicans (FICI = 0.5) [3]
- Limitations: Fer-1 has poor water solubility (requires DMSO dissolution) and is mainly used in preclinical studies; it has not been evaluated in clinical trials and has not been approved by the FDA for therapeutic use [2,4,6,7]

C15H22N2O2

262.35

262.168

C, 68.67; H, 8.45; N, 10.68; O, 12.20

347174-05-4

4068248

Gray to gray purple solid

1.1±0.1 g/cm3

437.3±35.0 °C at 760 mmHg

218.3±25.9 °C

0.0±1.1 mmHg at 25°C

1.595

3.9

2

4

5

19

290

0

O(C([H])([H])C([H])([H])[H])C(C1C([H])=C([H])C(=C(C=1[H])N([H])[H])N([H])C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C1([H])[H])=O

UJHBVMHOBZBWMX-UHFFFAOYSA-N

InChI=1S/C15H22N2O2/c1-2-19-15(18)11-8-9-14(13(16)10-11)17-12-6-4-3-5-7-12/h8-10,12,17H,2-7,16H2,1H3

3-amino-4-(cyclohexylamino)-benzoic acid, ethyl ester

Frer-1; 3-amino-4-(cyclohexylamino)-benzoic acid, ethyl ester; Ferrostatin-1; 347174-05-4; Ethyl 3-amino-4-(cyclohexylamino)benzoate; Fer-1; Ferrostatin-1 (Fer-1); Ferrostatin 1; ferrrostatin 1; MFCD08072959;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.5 mg/mL (9.53 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

---

Solubility in Formulation 2: 2.5 mg/mL (9.53 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

---

View More

Solubility in Formulation 3: ≥ 2.08 mg/mL (7.93 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

---

Solubility in Formulation 4: 0.2 mg/mL (0.76 mM) in 10% DMSO + 90% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

---

Solubility in Formulation 5: 2% DMSO+50% PEG 300+5% Tween 80+ddH2O: 5mg/mL

---

 (Please use freshly prepared in vivo formulations for optimal results.)