Hy 100579

Manufactured by MedChemExpress

Sourced in United States, China

HY-100579 is a laboratory equipment product manufactured by MedChemExpress. It is a versatile tool designed for various scientific applications. The core function of this product is to facilitate efficient and accurate measurements and manipulations required in laboratory settings.

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

13 protocols using hy 100579

Hypoxia-Induced Ferroptosis Regulation

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HTR-8/SVneo cells were obtained from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. Cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. Cells were grown at 37 °C, 5% CO₂.
The si-PPARγ, si-RXRα, si-Nrf2, oe-GPX4, and negative controls (si-NC and oe-NC) were purchased from HonorGene (Changsha, China). The si-PPARγ, si-RXRα, si-Nrf2, oe-GPX4, si-SREBP1, and negative controls were transfected into cells by Lipofectamine 2000. Rosiglitazone (122320-73-4, MedChemExpress, USA) is an agonist of PPARγ. Erastin (HY-15763, MedChemExpress, USA) is an agonist of ferroptosis and ferrostatin-1 (ferr1, HY-100579, MedChemExpress, USA) is an inhibitor. After treatment with 1 μM Rosiglitazone, 1 μM ferr1, and 1 μmol erastin for 24 h, cells were subjected to hypoxia for 24 h. As described earlier^{25 (link)}, the cells were placed under 2% O₂ for 24 h to establish a hypoxic cell model. Cells in the NC group were given normoxic conditions with 20% O₂. Cells were observed by a transmission electron microscope (jem-1400).

Lai W., Yu L, & Deng Y. (2024). PPARγ alleviates preeclampsia development by regulating lipid metabolism and ferroptosis. Communications Biology, 7, 429.

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Gef-resistant TNBC cell lines and ferroptosis

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Human TNBC cell lines MDA-MB-231 (ATCC^® CRM-HTB-26™) and HS578T (ATCC^® HTB-126™) (ATCC, VA, USA) were cultured in Dulbecco’s modified eagle’s medium (DMEM, 30-2002, ATCC) at 37℃ with 10% fetal bovine serum (FBS, 16000044, GIBCO, NY, USA). In order to construct Gef-resistant cells, MDA-MB-231 and HS578T cells were cultured in the medium with increasing concentration of Gefitinib for 11 months [the highest concentration of Gefitinib was 3 μM (21 )]. Finally, the Gef-resistant cells MDA-MB-231/Gef and HS578T/Gef were stored in the medium containing 3 μM Gefitinib (22 ). Gefitinib was purchased from MedChemExpress (HY-50895, Shanghai, China).
The Gef-resistant cells were assigned into sh-NC group (treated with lentivirus small hairpin RNA-negative control), sh-GPX4 group (treated with lentivirus sh-GPX4), sh-NC + DMSO group (treated with sh-NC + dimethyl sulphoxide), sh-GPX4 + DMSO (treated with lentivirus sh-GPX4 + DMSO), and sh-GPX4 + Ferrostatin-1 group (treated with lentivirus sh-GPX4 and 1 μM Ferrostatin-1) (23 (link)). Ferrostatin-1 is an inhibitor of ferroptosis and purchased from MedChemExpress (HY-100579). The titer of lentivirus was 1× 10⁸ Tu/ml.

Song X., Wang X., Liu Z, & Yu Z. (2020). Role of GPX4-Mediated Ferroptosis in the Sensitivity of Triple Negative Breast Cancer Cells to Gefitinib. Frontiers in Oncology, 10, 597434.

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Trophoblast Cell Line Manipulation

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The extravillous trophoblast cell line HTR-8/SVneo was purchased from the American Type Culture Collection and maintained in RPMI-1640 medium (HyClone; Cytiva) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin. All cells were cultured in a 37°C incubator with 5% CO₂. For BML-284 treatment, the cells were treated with 10 nM BML-284 (HY-19987; MedChemExpress) for 24 h after overexpression treatment for 48 h in a 37°C incubator with 5% CO₂. For ferrostatin-1 (Fer-1) treatment, cells were incubated with 1 µM ferrostatin-1 (HY-100579; MedChemExpress) for 24 h after overexpression treatment for 48 h in a 37°C incubator with 5% CO₂.

Li R., Weng X., Hu X., Wang J, & Zheng L. (2024). Pigment epithelium‑derived factor inhibits proliferation, invasion and angiogenesis, and induces ferroptosis of extravillous trophoblasts by targeting Wnt‑β‑catenin/VEGF signaling in placenta accreta spectrum. Molecular Medicine Reports, 29(5), 75.

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Investigating Ferroptosis and NADPH Oxidase in LPS-Induced Injury

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After dietary intervention, mice were subjected to various treatments. The mice were treated with a single dose of LPS (10 mg/kg, L2880, Sigma-Aldrich, Germany) or phosphate-buffered saline (PBS) via intraperitoneal injection.
For ferroptosis inhibition experiments, mice were injected intraperitoneally with ferrostatin-1 (10 mg/kg, HY-100579, MedChemExpress, USA) or vehicle 1 h before LPS administration. For NADPH oxidase inhibition, Vas2870 (10 mg/kg, HY-12804, MedChemExpress, USA) or vehicle was injected intraperitoneally 3 h before LPS administration. Twenty-four hours after LPS or PBS injection, mice were euthanized, and blood and kidneys were collected for further analysis.

Yao W., Liao H., Pang M., Pan L., Guan Y., Huang X., Hei Z., Luo C, & Ge M. (2022). Inhibition of the NADPH Oxidase Pathway Reduces Ferroptosis during Septic Renal Injury in Diabetic Mice. Oxidative Medicine and Cellular Longevity, 2022, 1193734.

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Establishing IDD Cell Model with tBHP

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NP tissues were harvested from the intervertebral discs of normal mice, cut into tiny pieces, detached with 0.25 mg/mL type II collagenase, and filtered (70 μm). The isolated NPCs were cultured in DMEM-F12 (Gibco) supplemented with 1% streptomycin-penicillin (Invitrogen) and 15% fetal bovine serum (FBS) (Gibco). NPCs were immunophenotypically characterized using anti-CD24 antibody (ab31622, Abcam) and anti-KRT18 antibody (ab215839, Abcam). Cells at passage 2 were considered suitable for subsequent experiments [20 (link)].
NPCs were exposed to different concentrations (25, 50, and 100 μM) of tert-butyl hydroperoxide (tBHP) (Sigma-Aldrich, St. Louis, MO) for 6 h or 12 h to establish an in vitro IDD cell model [21 (link)–24 (link)]. The ferroptosis-specific inhibitors ferrostatin-1 (5 μM, HY-100579, MedChemExpress, Monmouth Junction, NJ), liproxstatin-1 (5 μM, HY-12726, MedChemExpress), and deferoxamine (DFO) (100 μM, HY-B0988, MedChemExpress) were added to the NPC culture medium. EVs (1 μg/mL) were added to the NPC culture medium for coculture.

Yu X.J., Liu Q.K., Lu R., Wang S.X., Xu H.R., Wang Y.G., Bao Y., Jiang Y.Q., Li M.W, & Kang H. (2022). Bone Marrow Mesenchymal Stem Cell-Derived Extracellular Vesicles Carrying circ_0050205 Attenuate Intervertebral Disc Degeneration. Oxidative Medicine and Cellular Longevity, 2022, 8983667.

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Extracellular Vesicles Modulate Macrophage Function

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For determination of the effects of EVs on macrophages, EVs (10 μg/mL) isolated from T lymphocyte culture supernatants were cocultured with primary macrophages or RAW264.7 cells. EVs (20 μg/mL) isolated from human plasma were cocultured with THP-1 cells. After 24 h, the levels of various genes in RAW264.7 cells were evaluated by qPCR, and RAW264.7, primary macrophage or THP-1 cell lipid peroxidation and iron accumulation were detected. After 48 h, the expression of different proteins in RAW264.7 cells was measured by Western blots, and macrophage migration was measured by a Transwell experimental system. For inhibition of EV internalization, macrophages or RAW264.7 cells were pretreated with 20 μmol/L Dynasore (HY-15304, MedChemExpress, USA) for 1 h. For inhibition of lipid peroxidation, cells were pretreated with ferropstatin-1 (Fer-1, 5 μmol/L) (HY-100579, MedChemExpress, USA) or liproxstatin-1 (Lip-1, 1 μmol/L) (SML1414, Sigma-Aldrich, St. Louis, MO, USA) for 1 h. For prevention of free iron from engaging in chemical reactions, RAW264.7 cells were pretreated with the iron chelating agent deferoxamine mesylate (DFOM) (10 μmol/L) (HY–B0988, MedChemExpress, USA), while THP-1 cells were pretreated with DFOM (20 μmol/L) for 2 h.

Dang G., Li T., Yang D., Yang G., Du X., Yang J., Miao Y., Han L., Ma X., Song Y., Liu B., Li X., Wang X, & Feng J. (2022). T lymphocyte-derived extracellular vesicles aggravate abdominal aortic aneurysm by promoting macrophage lipid peroxidation and migration via pyruvate kinase muscle isozyme 2. Redox Biology, 50, 102257.

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Sepsis-Induced Ferroptosis and NETs Inhibition

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The ferroptosis inhibitor group was treated with ferrostatin-1 (1 mg/kg, Med Chem Express, HY-100579, Shanghai, China) immediately after CLP modeling. The NETs inhibitor group was treated with DNASE-1 (100 μg/kg, Beijing Solarbio Science & Technology Co., Ltd, 9003-98-9, Beijing, China) 1 h after modeling. The MEK/ERK pathway inhibition group was treated with U0126 (100 μg/kg, ABSIN, ABS810003, Shanghai, China) immediately after modeling.

Wang T., Zhang Z., Deng Z., Zeng W., Gao Y., Hei Z, & Yuan D. (2024). Mesenchymal stem cells alleviate sepsis-induced acute lung injury by blocking neutrophil extracellular traps formation and inhibiting ferroptosis in rats. PeerJ, 12, e16748.

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Cell line characterization and ferroptosis inhibition

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The human colon cancer cell lines (HT‐29, HCT‐116, SW480, SW620, LoVo, SW48, DLD‐1, Caco‐2, HCT‐15) and a normal human colon mucosal epithelial cell line NCM460 were purchased from the Chinese Academy of Sciences, Shanghai Institutes for Biological Sciences (Shanghai, China) and verified by short tandem repeat genotyping. Dulbecco's Modified Eagle Medium (DMEM, 11960077, Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (10099141, Gibco) was used to maintain all the cell lines under standard conditions (5% CO₂, 37°C). For the ferroptosis inhibition groups, ferrostatin‐1 (2 μmol/L, HY‐100579, MedChemExpress, Princeton, NJ, USA) was added to the complete culture medium.

Zheng X., Wang Q., Zhou Y., Zhang D., Geng Y., Hu W., Wu C., Shi Y, & Jiang J. (2022). N‐acetyltransferase 10 promotes colon cancer progression by inhibiting ferroptosis through N4‐acetylation and stabilization of ferroptosis suppressor protein 1 (FSP1) mRNA. Cancer Communications, 42(12), 1347-1366.

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Sorafenib and Ferrostatin-1 Induce Mitochondrial Morphology Changes

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After treatment with sorafenib (HY-10201, MedChemExpress, NJ, USA), ferrostatin-1 (Fer-1, HY-100579, MedChemExpress, NJ, USA) or DMSO for 24 h, the cells were fixed in 4% paraformaldehyde (P0099, Beyotime, Shanghai, China) for 2 min. Following cell harvest by centrifugation, 2.5% glutaraldehyde (P1126; Solarbio, Beijing, China) was carefully added to the cell pellet along the tube wall. Finally, ultrathin sections were cut, and the morphological changes of mitochondria were observed under TEM (TECNA I20; Philips, Eindhoven, Netherlands).

Xu X., Li Y., Wu Y., Wang M., Lu Y., Fang Z., Wang H, & Li Y. (2022). Increased ATF2 expression predicts poor prognosis and inhibits sorafenib-induced ferroptosis in gastric cancer. Redox Biology, 59, 102564.

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Cytotoxicity Assay of Cisplatin and Inhibitors

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Cells were seeded at a density of 5000 cells per well in 96-well plates (Corning Life Sciences, USA), grown overnight, and exposed to different concentrations of cisplatin (HY-17,394, MedChemExpress) and treated with different inhibitors, including 2-deoxy-D-glucose (HY-13,966, MedChemExpress), metformin (PHR1084, Merck), CB-839 (HY-12,248, MedChemExpress), liproxstatin-1 (HY-12,726, MedChemExpress) and ferrostatin-1 (HY-100,579, MedChemExpress), for 24 h, 48 h, 72 h, followed by 4 h of incubation with 10 µl of CCK8 reagent (Dojindo, Shanghai, China). The absorbance at 450 nm was measured using a microplate reader (Bio–Rad, USA).

Guo D., Zhang S., Gao Y., Shi J., Wang X., Zhang Z., Zhang Y., Wang Y., Zhao K., Li M., Wang A., Wang P., Gou Y., Zhang M., Liu M., Zhang Y., Chen R., Sun J., Wang S., Wu X., Liang Z., Chen J, & Lang J. (2023). Exploring the cellular and molecular differences between ovarian clear cell carcinoma and high-grade serous carcinoma using single-cell RNA sequencing and GEO gene expression signatures. Cell & Bioscience, 13, 139.

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