Ferrostatin 1

Manufactured by MedChemExpress

Sourced in United States, China

Ferrostatin-1 is a small molecule inhibitor that functions as a potent and selective inhibitor of ferroptosis, a form of regulated cell death driven by iron-dependent lipid peroxidation. It acts by blocking lipid peroxidation and ferroptosis without affecting other cell death pathways.

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Lab products found in correlation

Erastin, by MedChemExpress (21 mentions) Z vad fmk, by MedChemExpress (13 mentions) Necrostatin 1, by MedChemExpress (8 mentions) Necrosulfonamide, by MedChemExpress (7 mentions) Deferoxamine, by MedChemExpress (5 mentions) Foetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Cycloheximide (chx), by MedChemExpress (4 mentions) Sulfasalazine, by MedChemExpress (4 mentions) Sorafenib, by MedChemExpress (3 mentions) Bafilomycin a1, by Selleck Chemicals (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Hy 100579, by MedChemExpress (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Aicar, by MedChemExpress (3 mentions) Cisplatin, by Merck Group (2 mentions) Cck 8 kit, by Dojindo Laboratories (2 mentions) Fetal bovine serum (fbs), by Lonsera (2 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions) N acetyl l cysteine nac, by Beyotime (2 mentions)

Spelling variants of this product

Ferrostatin-1 (Fer-1) (22 citations) Ferrostatin-1 (HY-100579) (9 citations) Ferrostatin-1 (Fer-1; HY‐100579, (6 citations)

58 protocols using ferrostatin 1

Antibody-based Protein Detection and Inhibitor Assessment

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The antibodies used targeted the following proteins (dilutions used are included):
GAPDH (CW0101: immunoblotting, 1:1000) from CWBIOTECH; F-actin (40734ES75: immunofluorescence, 1:100) from YEASEN; U1A (10212: immunoblotting, 1:500) from Proteintech; CD63 (67605-1-Ig: immunoblotting, 1:5000) from Proteintech; CD9 (60232-1-Ig: immunoblotting, 1:5000) from Proteintech; TSG101 (14497-1-AP: immunoblotting, 1:1000) from Proteintech; GPX4 (67763-1-Ig: immunoblotting, 1:5000; IHC, 1:1000) from Proteintech; and DHODH (14877-1-AP: immunoblotting, 1:2000; IHC, 1:100) from Proteintech.
The inhibitors used are as follows:
Sorafenib (HY-10201: 10 μM for the in vitro assay and 30 mg/kg for the in vivo assay) and Ferrostatin-1 (HY-100579: 60 nM for the in vitro assay), and Ferrostatin-1 (HY-100579: 5 mg/kg, intraperitoneal injection for the in vivo assay) were obtained from MedChem Express.

Li X., Yu Q., Zhao R., Guo X., Liu C., Zhang K., Zhang W., Liu J., Yu J., Wang S., Hao Q., Li W., Zhang W., Li M., Zhang Y., Zhang C, & Gao Y. (2022). Designer Exosomes for Targeted Delivery of a Novel Therapeutic Cargo to Enhance Sorafenib-Mediated Ferroptosis in Hepatocellular Carcinoma. Frontiers in Oncology, 12, 898156.

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Evaluating Oxidative Stress in Pancreatic Cancer

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Pancreatic cancer cells were incubated for 72 h in 60 mm
² culture dishes with glutamine-free medium, medium containing 2 mM glutamine, or medium containing 2 mM glutamine supplemented with 50 μM diazooxonorleucine (Cat. #HY-108357; MedChemExpress, Shanghai, China) or 10 μM azaserine (Aza; Cat. #HY-B0919; MedChemExpress). Then, the cells were incubated with 10 μM 2’,7’-DCFH-DA (Cat. #D6883; Sigma-Aldrich, St Louis, USA) in serum-free DMEM at 37°C for 30 min. Cells treated with 100 μM H
₂O
₂ were used as positive controls. Thereafter, cells were treated with glutamine-free medium containing 2 μM ferrostatin-1 (Cat. #HY-100579; MedChemExpress), 5 mM N-acetylcysteine (NAC, Cat. #HY-B0215; MedChemExpress), 50 μM Z-VAD-FMK (Cat. #HY-16658B; MedChemExpress), and 150 μM necrostatin-1 (Cat. #HY-15760; MedChemExpress) to counteract cell death. The cells were then washed twice, resuspended, and filtered into single-cell suspensions before analysis. The fluorescence intensity of DCFH, formed by the reaction of DCFH-DA dyes with ROS, was measured at excitation and emission wavelengths of 488 and 530 nm, respectively, with a FC500 MPL flow cytometer (Beckman Coulter, Pasadena, USA). A minimum of 10,000 cells were analyzed per condition
[13] (link). Proportion analysis and histogram generation were performed using FlowJo software (version 10.6; FlowJo, Ashland, USA).

Xiao Z., Deng S., Liu H., Wang R., Liu Y., Dai Z., Gu W., Ni Q., Yu X., Liu C, & Luo G. (2023). Glutamine deprivation induces ferroptosis in pancreatic cancer cells: Glutamine deprivation induces pancreatic cancer ferroptosis. Acta Biochimica et Biophysica Sinica, 55(8), 1288-1300.

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Comprehensive Compound Acquisition Protocol

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Sorafenib (#T0093L) was purchased from TargetMol (Shanghai, China); lenvatinib (#S1164), brivanib (#S1084) and bafilomycin A1 (#S1413) were purchased from Selleck (Shanghai, China); cisplatin (#P4394), doxorubicin (#D1515), N-acetyl-l-cysteine (NAC, #A7250), and Hoechst (#94403) were purchased from Sigma‒Aldrich (Shanghai, China); canagliflozin (#A11100) was purchased from AdooQ Bioscience (Nanjing, China); and oligomycin (#HY-N6782), antimycin A (#HY-105755), phloretin (#HY-N0142), z-VAD-FMK (#HY-16658B), necrostatin-1 (#HY-15760), and ferrostatin-1 (#HY-100579) were purchased from MedChemExpress (Shanghai, China).

Zhou J., Feng J., Wu Y., Dai H.Q., Zhu G.Z., Chen P.H., Wang L.M., Lu G., Liao X.W., Lu P.Z., Su W.J., Hooi S.C., Ye X.P., Shen H.M., Peng T, & Lu G.D. (2022). Simultaneous treatment with sorafenib and glucose restriction inhibits hepatocellular carcinoma in vitro and in vivo by impairing SIAH1-mediated mitophagy. Experimental & Molecular Medicine, 54(11), 2007-2021.

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Sorafenib and Ferrostatin-1 Protocol

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Sorafenib (#HY-10201) and Ferrostatin-1 (#HY-100579) were purchased from MedChemExpress (New Jersey, United States).

Li Y., Yan J., Zhao Q., Zhang Y, & Zhang Y. (2022). ATF3 promotes ferroptosis in sorafenib-induced cardiotoxicity by suppressing Slc7a11 expression. Frontiers in Pharmacology, 13, 904314.

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Ferroptosis Molecular Assays Protocol

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Prussian-blue kit (Solarbio, Beijing, China), ROS Assay kit (Beyotime, Shanghai, China), Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme, Nanjing, China), CCK-8 kit, and Liperfluo (Dojindo, Shanghai, China) were used following instructions. All ELISA kits were purchased from Dakewe (BioLegend, California, USA). Ferrostatin-1 (HY-100579), Liproxtatin-1 (HY-12726) and RSL3 (HY-100218A) was purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA) and prepared in DMSO. M-CSF (Novoprotein, Shanghai, China) for BMDM was prepared in ddH₂O. NIR-797 and Rhodamine B (Hualanchem, Shanghai, China) were prepared in DMSO. The antibodies used for flow cytometry (supplemental Table S1) and PCR Assay plates (Wcgene Biotech, Shanghai, China) were all purchased from fcmacs (Nanjing, China). SPIO nanoparticles (Ferumoxytol) were kindly provided by professor Ning Gu from Southeast University.

Pan Y., Li J., Wang J., Jiang Q., Yang J., Dou H., Liang H., Li K, & Hou Y. (2022). Ferroptotic MSCs protect mice against sepsis via promoting macrophage efferocytosis. Cell Death & Disease, 13(9), 825.

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Culturing Liver Cancer Cell Lines

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Human liver cancer cells HepG2 were obtained from the American Type Culture Collection (ATCC). HCCC9810 was obtained from Procell (Catalog, CL-0095). Cell cultures were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Grand Island, NY, USA) with 10% fetal bovine serum (FBS) (EallBio, Beijing, China; # 03. U16001) at 37°C in a humidified atmosphere of 95% air and 5% CO₂. RSL3, dimethyl sulfoxide (DMSO), Z-VAD-FMK, necrosulfonamide, and ferrostatin-1 were purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA).

Cao F., Hao W., Liang W., Zeng H, & Zheng J. (2024). MiR-339-5p Inhibits Ferroptosis by Promoting Autophagic Degradation of FTH1 Through Targeting ATG7 in Liver Cancer Cells. Clinical Medicine Insights. Oncology, 18, 11795549241244783.

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Macrophage Ferroptosis Induction Assay

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Macrophages were implanted in a 96-well plate and cultured for 24 h at 37 °C. After macrophages had undergone different treatments at specific times, the supernatant was removed, and then the cells were washed with PBS three times. 100 μL 1640 medium containing 10 μL Cell Counting Kit-8 (CCK-8, Byotime, Beijing, China) was added into each well. The cells were incubated for 1 h at 37 °C until they turned orange and then measured at an absorbance of 450 nm. All drugs and corresponding treatment concentration: erastin (HY-15763), 10 μM; RSL3 (HY-100218A), 5 μM Ferrostatin-1 (HY-100579), 10 μM; deferoxamine (HY-B0988), 200 μM; necrostatin (HY-15760) 2 μM; ZVAD-FMK(HY-16658B), 5 μM were purchased from MedChemExpress (MCE). FeSO₄ 7H2O (F7002), 200 μg/mL; baicalein (465119), 20 μM were purchased from Sigma.

Yi Z.H., Li S.Q., Ke J.Y., Wang Y., Zhao M.Z., Li J., Li M.Q, & Zhu Z.L. (2022). Baicalein Relieves Ferroptosis-Mediated Phagocytosis Inhibition of Macrophages in Ovarian Endometriosis. Current Issues in Molecular Biology, 44(12), 6189-6204.

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Osteoblastic Cell Responses to MaR1 in T2DM

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The osteoblastic cell line MC3T3-E1 was provided by the Cell Resource Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences. Fetal bovine serum (FBS; Lonsera, Salto, Uruguay)-supplemented α-minimum essential medium (α-MEM; Biological Industries, Acre, Israel) was used to cultivate cells at 37 °C in a humidified atmosphere with 5% CO₂. Cells were cultured in the appropriate osteogenic induction medium, which contained 10 nM dexamethasone, 50 mg/L ascorbic acids, and 10 mM β-glycerophosphate (Solarbio, Beijing, China).
MC3T3-E1 cells were cultured in various groups for 72 h to test the effects of MaR1, including Normal medium (α-MEM with 5.5 mM glucose), control medium (with the addition of 20 mM mannitol and palmitate vehicle to the Normal medium of 5.5 mM glucose; Sigma, Ronkonkoma, NY, USA), T2DM medium (25 mM glucose and 200 mM sodium palmitate to mimic diabetic conditions; Alladin, Shanghai, China), and T2DM medium with MaR1 (1 or 10 nM; Cayman, Ann Arbor, MI, USA). To investigate the existence and potential mechanism of ferroptosis in osteoblasts, the ferroptosis inhibitor ferrostatin-1 (Fer-1, 5 μM; MedChemExpress, Monmouth Junction, NJ, USA) and the ferroptosis activator Erastin (Era, 1 μM; MedChemExpress, Junction, NJ, USA) were administered to the cell cultures.

Zhang Z., Ji C., Wang Y.N., Liu S., Wang M., Xu X, & Zhang D. (2022). Maresin1 Suppresses High-Glucose-Induced Ferroptosis in Osteoblasts via NRF2 Activation in Type 2 Diabetic Osteoporosis. Cells, 11(16), 2560.

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Cell Viability Assay Using Resazurin

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Viability was determined by resazurin dye (Sigma-Aldrich, #R7017). Briefly, 3000 cells per well were seeded into 96-well plate at a final volume of 200 μL. Depending on the cell line, at day 4 or 6 resazurin was added to a final concentration of 30 μmol L⁻¹, and plates were left in the incubator for 3–5 h. Fluorescence (λ_ex = 525 nm; λ_em = 590 nm) was measured 3 h later and viability calculated as a percentage relative to the untreated well. Every experiment was performed by technical triplicate and repeated at least 3 times. Reagents used were: 1S,3S-(RSL3) (MedChem Express, #HY-100218A), l-buthionine sulfoximine L-BSO (Sigma, #B2515), Ferrostatin-1 (MedChem Express, # HY-100579), Necrostatin-1 (MedChem Express, # HY-15760), quinoline-Val-Asp-Difluorophenoxymethylketone (QVD) (MedChem Express, # HY-12305), tert-butyl hydroquinone tBHQ (MedChem Express, #HY-100489), 5′-aza-2′-deoxycytidine (DEC) (Sigma, # A3656). Fluorescence was determined in a Synergy H1 microplate reader (Biotek).

Pontel L.B., Bueno-Costa A., Morellato A.E., Carvalho Santos J., Roué G, & Esteller M. (2022). Acute lymphoblastic leukemia necessitates GSH-dependent ferroptosis defenses to overcome FSP1-epigenetic silencing. Redox Biology, 55, 102408.

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Ferroptosis Pathway Regulation Assay

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Reagents: ferrostatin‐1 (CAS:347174–05‐4) and erastin (CAS:571203–78‐6) were all purchased from MedChemExpress (shanghai; China). SBFI26 (CAS:1541207–06‐0) was purchased from GLPBIO Technology (American). Antibodies: Primary antibodies were used as following: anti‐β‐actin antibody (1:1000, mAbcam 8226, Abcam), anti‐ALOX5 (1:500–1:1000, R1512‐14, HuaBio), anti‐ALOX12 (1:2000, AP8877B Abcepta Biotech), anti‐NFE2L2 (1:1000–1:2000, R1312‐8, HuaBio), anti‐GPX4 (1:500–1:2000, ET1706‐45, HuaBio), anti‐HO‐1 (1:1000, ET1604‐45, HuaBio), anti‐ATF4 (1:500–1:2000, ET1612‐37, HuaBio), secondary antibody was purchased from Beyotime (Shanghai, China).

He G., Zhang Y., Feng Y., Chen T., Liu M., Zeng Y., Yin X., Qu S., Huang L., Ke Y., Liang L., Yan J, & Liu W. (2024). SBFI26 induces triple‐negative breast cancer cells ferroptosis via lipid peroxidation. Journal of Cellular and Molecular Medicine, 28(7), e18212.

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