Ml210

Manufactured by Merck Group

The ML210 is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It features a modular design, allowing users to configure the system based on their specific requirements. The ML210 is capable of delivering precise and accurate solvent flow rates, providing reliable and reproducible results for a wide range of chromatographic separations.

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Lab products found in correlation

Ferrostatin 1, by Merck Group (6 mentions) Erastin, by Merck Group (2 mentions) Bortezomib, by Thermo Fisher Scientific (2 mentions) Celltiter glo luminescent cell viability assay, by Promega (2 mentions) N acetyl l cysteine, by Merck Group (2 mentions) α tocopherol, by Merck Group (2 mentions) Nsc23766, by Merck Group (2 mentions) Mk571, by Bio-Techne (2 mentions) Quercetin, by Bio-Techne (2 mentions) Jzl184, by Cayman Chemical (2 mentions) Ml239, by Merck Group (2 mentions) Sc 26196, by Bio-Techne (2 mentions) Cp100356, by Merck Group (2 mentions) Obatoclax mesylate, by Selleck Chemicals (2 mentions) Ym155, by Selleck Chemicals (2 mentions) Elacridar, by Merck Group (2 mentions) Arachidonic acid, by Cayman Chemical (1 mentions) Linoleic acid, by Cayman Chemical (1 mentions) Oleic acid, by Cayman Chemical (1 mentions) Eicosapentaenoic acid, by Cayman Chemical (1 mentions)

9 protocols using ml210

Ferroptosis Induction and Inhibition Assays

Cited 2 times

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The following compounds were obtained from Sigma-Aldrich: ML210 (SML0521, purity ≥ 98%, used at wide concentration range), 1S, 3R-RSL3 (RSL3, SML2234, purity ≥ 98%, used at wide concentration range), ferrostatin-1 (Fer-1, SML0583, purity ≥ 95%, used at 5 μM), erastin (E7781, purity ≥ 98%, used at wide concentration range) and L-buthionine-[S,R]-sulfoximine (BSO, B2515, purity ≥ 97%, used at wide concentration range). Free fatty acids, including oleic acid (OA, C18:1), linoleic acid (LA, C18:2), a-linolenic acid (ALA, C18:3), arachidonic acid (AA, C20:4), eicosapentaenoic acid (EPA, C20:5), docosapentaenoic acid (DPA, C22:5), and docosahexaenoic acid (DHA, C22:6) were purchased from Cayman Chemicals and conjugated with fatty-acid free BSA (Sigma-Aldrich) using previously described protocols⁵¹, and treated cells at 20 μM for 3 days in the viability assays. The following compounds were prepared according to known literature procedures: FINO2^{36 (link)} and FIN56^{34 (link)}. Alkyne analogs of ML210 and RSL3 (ML210-yne and RSL3-yne, respectively) were synthesized as previously described^{15 (link)}.

Zou Y., Li H., Graham E.T., Deik A.A., Eaton J.K., Wang W., Sandoval-Gomez G., Clish C.B., Doench J.G, & Schreiber S.L. (2020). Cytochrome P450 oxidoreductase contributes to phospholipid peroxidation in ferroptosis. Nature chemical biology, 16(3), 302-309.

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High-throughput Screening of Ferroptosis Compounds

Cited 1 time

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ML162 was synthesized by Acme (Palo Alto, CA). Erastin2 (Cat# 27087) and deferoxamine (Cat# 14595) were obtained from Cayman Chemicals (Ann Arbor, MI). Palbociclib (Cat# S1116), pimasertib (Cat# S1475), and 1S,3R-RSL3 (simply, RSL3) (Cat# S8155) were obtained from Selleck Chemicals (Houston, TX). Doxycycline (Dox) (Cat# D3447), ML210 (Cat# SML0521), ferrostatin-1 (Cat# SML0583) and propidium iodide (Cat# P4170) were obtained from Sigma-Aldrich (St. Louis, MO). Abemaciclib (Cat# HY-16297A) was obtained from MedChem Express (Princeton, NJ). Bortezomib (Cat# NC0587961), Q-VD-OPh (Cat# OPH00101M), etoposide (Cat# 12-261-00), DAPI (Cat# D1306) and C11 BODIPY 581/591 were from Thermo Fisher Scientific (Waltham, MA). Compound 28 was synthesized as described⁴⁷ and provided as a gift by P. Beltran (Ferro Inc). Dox was dissolved in deionized water and stored at −20°C. C11 BODIPY 581/591 was dissolved into methanol and stored at −20°C. All other compounds were dissolved in dimethyl sulfoxide (DMSO) and then stored at −20°C. The compound library (Cat# L1700) was obtained from Selleck Chemicals^{8 (link)}.

Rodencal J., Kim N., Li V.L., He A., Lange M., He J., Tarangelo A., Schafer Z.T., Olzmann J.A., Sage J., Long J.Z, & Dixon S.J. (2023). A Cell Cycle-Dependent Ferroptosis Sensitivity Switch Governed by EMP2. bioRxiv.

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Ferroptosis Inducers and Inhibitors Protocol

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ML210 (SML0521), RSL3 (SML2234), ferrostatin-1 (SML0583), liproxstatin-1 (SML1414), erastin (E7781) and necrostatin-1 (N9037) were obtained from Sigma-Aldrich, all of which exhibit purity ≥98% by HPLC. z-VAD-FMK was purchased from Thermo Fisher Scientific (FMK001). AGPS inhibitor ZINC-69435460 was synthesized following previously described procedures^{13 (link)}. FIN56 was synthesized as previously described (#WO2008140792A1)^{32 ,33}.

Zou Y., Henry W.S., Ricq E.L., Graham E.T., Phadnis V.V., Maretich P., Paradkar S., Boehnke N., Deik A.A., Reinhardt F., Eaton J.K., Ferguson B., Wang W., Fairman J., Keys H.R., Dančík V., Clish C.B., Clemons P.A., Hammond P.T., Boyer L.A., Weinberg R.A, & Schreiber S.L. (2020). Plasticity of ether lipids promotes ferroptosis susceptibility and evasion. Nature, 585(7826), 603-608.

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Assessing Ferroptosis Sensitivity in Cancer Cells

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Cells were seeded in 96-well black clear bottom plates (Corning) at 2000 or 3000 cells (1099 +/− TGF-β) and 6000 cells (OVCAR8) per well. Approximately, 12–16 h post-seeding, cells were treated with various drug concentrations using an HP D300e Digital Dispenser unless stated otherwise. Cell viability was assessed at 72 h post-treatment by performing CellTiter-Glo Luminescent Cell Viability Assays (Promega) according to the manufacturer’s instructions. Relative viability was calculated by normalizing to untreated controls unless stated otherwise. Non-linear regression models were applied to generate the regression fit curves using GraphPad Prism. Drug compounds were purchased as indicated: RSL3 (Selleck Chem), ML210 (Sigma Aldrich), and Liproxstatin-1 (Fisher Scientific). For experiments involving ferric ammonium citrate (FAC), FAC (Sigma) was prepared fresh in sterile 1× PBS and manually added directly to cell culture media at the indicated concentrations at the time of seeding into 96-well plates. Unless stated otherwise, cells were pretreated with FAC for 24 h prior to ML210 treatment.

Henry W.S., Müller S., Yang J.S., Innes-Gold S., Das S., Reinhardt F., Sigmund K., Phadnis V.V., Wan Z., Eaton E., Sampaio J.L., Bell G.W., Viravalli A., Hammond P.T., Kamm R.D., Cohen A.E., Boehnke N., Hsu V.W., Levental K.R., Rodriguez R, & Weinberg R.A. (2024). Ether lipids influence cancer cell fate by modulating iron uptake. bioRxiv.

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Ferroptosis Induction and Measurement Protocol

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Erastin2 was synthesized by Acme Bioscience (Palo Alto, CA). RSL3 (Cat# S8155) and INK128 (Cat# S2811) were from Selleck Chemicals (Houston, TX). SYTOX Green (Cat# S7020) was from Life Technologies. Dimethyl sulfoxide (DMSO; Cat# 276855), ferrostatin-1 (Cat# SML0583), ML210 (Cat# SML0521), methanol (Cat# 34860), rosiglitazone (Cat# R2408), staurosporine (Cat# S6942), thapsigargin (Cat# T9033), and vinblastine (Cat# V1377) were from Sigma-Aldrich (St. Louis, MO). Bortezomib (Cat# NC0587961) and camptothecin (Cat# AC276721000) were from Fisher Scientific. ZINC-69435460 (AGPSi; Cat# Z1030248250) was from Enamine US Inc. (Cincinnati, OH). C11 BODIPY 581/591 (Cat# D3861) and Hoechst (Cat# H1399) were from Molecular Probes (Eugene, OR). FINO₂ was a gift from Keith Woerpel (New York University, NY, NY). CIL56 and FIN56 were gifts from Rachid Skouta (UMass Amherst, Amherst, MA). C11 BODIPY 581/591 was dissolved in anhydrous methanol, and all other chemicals were dissolved in DMSO. All chemicals were stored at −20°C until use.

Magtanong L., Mueller G.D., Williams K.J., Billmann M., Chan K., Armenta D.A., Moffat J., Boone C., Myers C.L., Olzmann J.A., Bensinger S.J, & Dixon S.J. (2022). Context-Dependent Regulation of Ferroptosis Sensitivity. Cell chemical biology.

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Small Molecule Acquisition and Preparation

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JZL184 (Cat. no. 13158; ≥97% purity) was purchased from Cayman Chemical. ML239 (SML0442; ≥98%), NSC23766 (SML0952; ≥97%), CP-100356 (PZ0171; ≥98%), elacridar (SML0486; ≥98%), ML210 (SML0521; ≥98%), ferrostatin-1 (SML0583; ≥95%), α-tocopherol (T3251; ≥99%), and N-acetyl-L-cysteine (A7250; ≥99%) were purchased from Sigma. YM-155 (S1130; ≥99%), obatoclax mesylate (S1057; ≥99%), and RITA (S2781; 99%) were purchased from SelleckChem. SC-26196 (4189; 99.2%), quercetin (1125; >98%), and MK-571 (2238; >96.9%) were purchased from Tocris. Austocystin D was purchased from eMolecules. The selective CYP2J2 inhibitor 1-(4-bromophenyl)-4-[4-(hydroxydiphenylmethyl)piperidin-1-yl]butan-1-one was synthesized in-house according to the published methodology (>95% purity)^{48 (link)}. All small molecules were dissolved in DMSO, except N-acetylcysteine, which was dissolved in cell culture medium then adjusted to pH 7.5.

Rees M.G., Seashore-Ludlow B., Cheah J.H., Adams D.J., Price E.V., Gill S., Javaid S., Coletti M.E., Jones V.L., Bodycombe N.E., Soule C.K., Alexander B., Li A., Montgomery P., Kotz J.D., Hon C.S., Munoz B., Liefeld T., Dančík V., Haber D.A., Clish C.B., Bittker J.A., Palmer M., Wagner B.K., Clemons P.A., Shamji A.F, & Schreiber S.L. (2015). Correlating chemical sensitivity and basal gene expression reveals mechanism of action. Nature chemical biology, 12(2), 109-116.

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Small Molecule Acquisition and Preparation

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Reagent Sources for Cell Experiments

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RSL3 was purchased from Fisher Scientific (Cat# 611810). ML210 (Cat#
SML0521), cisplatin (Cat# 1134357), and carboplatin (Cat# C2538) were from
Sigma-Aldrich. WNT3a was from Fisher Scientific (Cat# 5036WN010CF, R&D
Systems), and IWR-1-endo was from Santa Cruz (sc-295215A).

Wang Y., Zhao G., Condello S., Huang H., Cardenas H., Tanner E.J., Wei J., Ji Y., Li J., Tan Y., Davuluri R.V., Peter M.E., Cheng J.X, & Matei D. (2020). Frizzled-7 identifies platinum-tolerant ovarian cancer cells susceptible to ferroptosis. Cancer research, 81(2), 384-399.

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Cell Viability Assay with Small Molecules

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For cell viability experiments, cells were treated with a range of concentrations of the indicated small molecule compounds, and relative viability was measured by the CellTiter-Glo Assay. Cells were seeded in 96-well black-wall tissue culture-treated plates at 2,000 to 3,000 cells per well (or 10,500 for primary human monocytes and macrophages), to reach 30% confluency the following day. Cells were treated with compounds 18–24 h after seeding using an HP D300e Digital Dispenser with three biological replicates per condition. For BSA-palmitate and BSA-control experiments, compounds were added manually to cells. 66–72 h after compound treatment, viability was measured using the CellTiter-Glo Luminescent Cell Viability Assay (Promega) as per the manufacturer’s instructions. Unless otherwise specified, relative viability was normalized to the untreated condition. Regression fit curves were computed in Prism 8 or 9 (GraphPad) using four-parameter inhibition nonlinear regression. The mean and standard deviation for three biological replicates of each data point were calculated. The following compounds were used: RSL3 (Selleck Chem), ML210 (Sigma Aldrich), erastin (Selleck Chem), FIN56 (Selleck Chem), Ferrostatin-1 (Sigma Aldrich), Z-VAD-FMK (Selleck Chem), Necrostatin-1 (Selleck Chem), BSA-palmitate (Cayman Chem), BSA-control (Cayman Chem).

Mahr K., Hillen W, & Titgemeyer F. (2000). Carbon Catabolite Repression in Lactobacillus pentosus: Analysis of the ccpA Region. Applied and Environmental Microbiology, 66(1), 277-283.

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