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Ethacrynic acid

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Ethacrynic acid is a chemical compound used in laboratory settings. It is a potent diuretic agent that inhibits sodium reabsorption in the kidneys. Ethacrynic acid is commonly used in research applications as a pharmacological tool to study renal function and electrolyte balance.

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Lab products found in correlation

Jy92 iin, by Ningbo Scientz Biotechnology (3 mentions) Dansylcadaverine, by Merck Group (1 mentions) Coomassie brilliant blue, by Thermo Fisher Scientific (1 mentions) Aptt reagent, by Thermo Fisher Scientific (1 mentions) Schrodinger suite 2020, by Schrödinger (1 mentions) N n dimethylcasein, by Merck Group (1 mentions) Dithiothreitol (dtt), by Merck Group (1 mentions) Fibrinogen, by Prolytix (1 mentions) Fxiiia, by Prolytix (1 mentions) α2 antiplasmin, by Prolytix (1 mentions) Hematin, by Merck Group (1 mentions) Bromosulfophthalein, by Merck Group (1 mentions) Phenylmethanesulfonyl fluoride, by Merck Group (1 mentions) 1 2 dichloro 4 nitrobenzene, by Merck Group (1 mentions) Oxidized glutathione, by Merck Group (1 mentions) Reduced glutathione, by Merck Group (1 mentions) Melphalan, by Merck Group (1 mentions) Posaconazole, by Merck Group (1 mentions) Ifosfamide, by Merck Group (1 mentions) Levetiracetam, by Merck Group (1 mentions)

Close spelling variants of this product

Ethacrynic acid (EA; (2 citations) Ethacrynic acid (ECA) (2 citations)

11 protocols using ethacrynic acid

1

Anticoagulant Effects on Factor XIII Activity

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Ethacrynic acid was obtained from Sigma Aldrich (St. Louis, MO). Human plasma thrombin, α2-antiplasmin, and FXIIIa were obtained from Haematologic Technologies (Essex Junction, VT). N,N-dimethyl-casein, dansyl-cadaverine, and dithiothreitol (DTT) were also from Sigma Aldrich. Dabigatran, rivaroxaban, and AntiF11 (mouse monoclonal antibody from Abnova™) for plasma studies as well as Coomassie Brilliant Blue for gel electrophoresis were from Fisher Scientific (Waltham, MA). Fibrinogen was from Haematologic Technologies. Stock solution of FXIIIa was prepared in 50 mM Tris–HCl, 1 mM CaCl2, 100 mM NaCl, 0.1% PEG8000, 0.02% Tween80, and 2 mg/mL N,N–dimethylcasein. For the clotting assays, pooled normal human plasma was purchased from George King Bio-Medical (Overland Park, KS). APTT reagent containing ellagic acid, thromboplastin‒D (PT reagent), and 25 mM solution of CaCl2 were purchased from Thermo Fisher Scientific. All experiments in this paper were repeated at least two times. For molecular modeling studies, initial structure of FXIIIa (4kty.pdb) was prepared by removing the crystallographic water molecules and adding hydrogen atoms consistent with the physiologic pH of 7 using Maestro 12.4.1. Docking studies were carried out by generating a non-covalent pose using Glide (Schrodinger Suite 2020), and then, by using the covalent docking program (Schrodinger Suite 2020).
Kar S., Vu K., Mottamal M, & Al-Horani R.A. (2022). Ethacrynic acid is an inhibitor of human factor XIIIa. BMC Pharmacology & Toxicology, 23, 35.
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2

Glutathione-Conjugating Enzyme Assay Protocol

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Reduced glutathione (GSH), oxidized glutathione (GSSG), 1-chloro 2,4-dintrobenzene (CDNB), 7-chloro-4-nitrobenzene-2-oxa-1,3-diazole, ethacrynic acid, 1,2-dichloro-4-nitrobenzene (DCNB), bromosulfophthalein (BSF), ρ-nitrophenyl acetate (ρNPA), phenylmethanesulfonyl fluoride, cibracron blue, hematin are products of Sigma-Aldrich Chemical, St Louis, USA. DEAE-Trisacryl was from LKB, Villeneuve-la Garenne, France. Tributyltinacetate (TBTA) and triphenyltinchloride (TPTC) were purchased from Alfa Company (Karlerushe, Germany). Glutathione Sepahrose 4B™ is from Amersham Bioscience, Uppsala, Sweden. Other specialty chemicals were obtained from Sigma in the highest purity available unless noted otherwise. All other reagents were of analytical grade commercially available.
, & Kolawole A.O. (2016). Catalysis of Silver catfish Major Hepatic Glutathione Transferase proceeds via rapid equilibrium sequential random Mechanism. Toxicology Reports, 3, 598-607.
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3

Cytotoxic Agents and Cell Lines

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The J45.01 T-cell line and Toledo B-cell line were obtained from American Type Culture Collection (Manassas, VA). KBM3/Bu2506 is a busulfan-resistant AML cell line established in our laboratory as previously described [10 (link)]. MOLM14 is an AML cell line obtained from Dr. Michael Andreeff's laboratory (UT MD Anderson Cancer Center, Houston, TX). All cells were cultured in RPMI 1640 (Mediatech, Manassas, VA) supplemented with 10% heat-inactivated fetal bovine serum (FBS: Sigma-Aldrich, St. Louis, MO) and 100 U/mL penicillin and 100 μg/mL streptomycin (Mediatech) at 37°C in a humidified atmosphere of 5% CO2 in air. 5-Carboxyfluorescein diacetate acetoxymethyl ester (5-CFDA) was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA). Verapamil and MK571 were purchased from Selleckchem (Houston, TX). Chlorambucil, busulfan, melphalan, bendamustine, cyclophosphamide, ifosfamide, ketoconazole, posaconazole, fluconazole, itraconazole, metronidazole, ethacrynic acid, everolimus, sirolimus, phenytoin, levetiracetam, and buthionine sulphoximine (BSO) were obtained from Sigma-Aldrich Corp. (St. Louis, MO). 4-Hydroperoxycyclophosphamide (4-HC) and 4-hydroperoxyifosfamide (4-HI) were generous gifts from from Dr. Scott Rowley (Hackensack University Medical Center, Hackensack, New Jersey), and Dr. Robert F. Struck (Southern Research Institute, Birmingham, Alabama) respectively.
Valdez B.C., Hassan M, & Andersson B.S. (2017). Development of an assay for cellular efflux of pharmaceutically active agents and its relevance to understanding drug interactions. Experimental hematology, 52, 65-71.
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4

Multimodal Nanodrug Delivery Protocols

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O2‐(2,4‐Dinitrophenyl) 1‐[(4‐ethoxycarbonyl) piperazin‐1‐yl] diazen‐1‐ium‐1,2‐diolate (JS‐K, molecular weight (Mw = 384.30)), (2S,3R,4E)‐2‐Amino‐4‐octadecene‐1,3‐diol 1‐phosphate, D‐erythro‐Sphingosine 1‐phosphate (Sphingosine‐1‐phosphate, S1P), 2‐[2,3‐Dichloro‐4‐(2‐methylene‐1‐oxobutyl)phenoxy]acetic acid (Ethacrynic acid, ECA) and 4‐methyl‐5‐oxo‐2,3,4,6,8‐pentazabicyclo [4.3.0] nona‐2,7,9‐triene‐9‐carboxamide (Temozolomide, TMZ) were all purchased from Sigma‐Aldrich (USA). 1,2‐dipalmitoyl‐sn‐glycero‐3‐phosphocholine (DPPC), 1,2‐Distearoylsn‐sn‐glycero‐3‐phosphocholine (DSPC), 1,2‐distearoyl‐sn‐glycero‐3‐phosphoethanolamine‐N‐[methoxy(poly(ethylene glycol))‐2000] (DSPE‐mPEG2000) were all purchased from Jiangsu Southeast Nanomaterials Co., Ltd (Huaian, China). 3,3′‐Dioctadecyloxacarbocyanine perchlorate (DiO), 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethylindotricarbocyanine iodide (DiR), and 1,1′‐Dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI) were purchased from Beyotime (Haimen, China). All chemicals were analytical grade and used without further purification.
Liu Y., Cui L., Wang X., Miao W., Ju Y., Chen T., Xu H., Gu N, & Yang F. (2023). In Situ Nitric Oxide Gas Nanogenerator Reprograms Glioma Immunosuppressive Microenvironment. Advanced Science, 10(18), 2300679.
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5

Breast Cancer Cell Line Cultivation

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All cells were obtained from the Institute of Biochemistry and Cell Biology of Chinese Academy of Science (Shanghai, China). Human breast cancer cell lines (MCF7 and MDA-MB-231) and mouse breast cancer cells (4T1) were maintained in RPMI 1640 with 10% fetal bovine serum (FBS) (PAA Laboratories, Pasching, Australia) in a humidified atmosphere containing 5% CO2 at 37°C. Afatinib and neratinib were purchased from Selleck Chemicals company (USA), and ethacrynic acid was obtained from Sigma (USA).
Liu B., Huang X., Hu Y., Chen T., Peng B., Gao N., Jin Z., Jia T., Zhang N., Wang Z, & Jin G. (2016). Ethacrynic acid improves the antitumor effects of irreversible epidermal growth factor receptor tyrosine kinase inhibitors in breast cancer. Oncotarget, 7(36), 58038-58050.
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6

Evaluating Drug Resistance in Cell Cultures

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For resistance determination, 40,000 cells were seeded per well in a clear bottom-flat 24-well plate (Greiner) including medium control as blank and were grown overnight. The cells were exposed to increasing concentrations from 0.2 to 500 μM SM including vehicle control for 5 days. Glutathione S-transferase (GST) inhibitors ezatiostat (TLK199, Cayman Chemical) and ethacrynic acid (Sigma) were added to medium in final concentrations of 400 nM and 600 nM, respectively. Stock solutions of both inhibitors were prepared in DMSO with 10 mg/mL. After pre-dilution in 100 % EtOH, GST activator ethoxyquin (Sigma) was used with 24 μM final concentration as a pre-treatment for 24 hours. For testing resistance against H2O2, concentrations between 0.24 and 80,000 μ M were used including medium control for 5 days. Therefore, 30 % H2O2 (Sigma) was pre-diluted in sterile ddH2O. After exposition over 5 days, cells were washed with medium and XTT staining solution (Sigma) was prepared by mixing 5 mL XTT labeling reagent with 0.1 mL electron-coupling reagent per plate. 400 μ L medium with 200 μ L of XTT staining solution were added per well and incubated for 6 h. Extinction was measured at 450 nm with reference at 630 nm. All conditions were tested in biological triplicates.
Rothmiller S., Schröder S., Strobelt R., Wolf M., Wang J., Jiang X., Worek F., Steinritz D., Thiermann H, & Schmidt A. (2017). Sulfur mustard resistant keratinocytes obtained elevated glutathione levels and other changes in the antioxidative defense mechanism. Toxicology letters, 293, 51-61.
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7

Glutathione S-Transferase Activity Assay

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After AgNPs-, AuNPs- or Ag+-treatment, cells were washed with PBS for 3 times, and harvested by centrifugation at 1000 rpm. Then, cell pellets were resuspended in cold buffer of a Glutathione S- transferase Assay Kit (Solarbio Science & Technology Co., Ltd., Beijing, China) and treated with a ultrasonic homogenizer (JY92-IIN, Ningbo Scientz Biotechnology Co.,Ltd. China) for 3 min in ice bath. After centrifugation at 12,000 g for 10 min at 4 °C, the supernatant was used for enzyme activity measurement according to the manufacturer’s instructions. For GST activity inhibition, ethacrynic acid (≥97%, Sigma-Aldrich, USA) was dissolved in dimethyl sulfoxide as the stock solution of 10 mg/mL. A commercial GST purified from equine liver (≥ 25 units/mg protein, Sigma-Aldrich, USA) was dissolved in PBS as the stock solution of 5 mg/mL.
Xu M., Yang Q., Xu L., Rao Z., Cao D., Gao M, & Liu S. (2019). Protein target identification and toxicological mechanism investigation of silver nanoparticles-induced hepatotoxicity by integrating proteomic and metallomic strategies. Particle and Fibre Toxicology, 16, 46.
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8

Edaravone's Cytoprotective Effects on TDP-43 Mutants

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Edaravone synthesized at Mitsubishi Tanabe Pharma Corporation (Osaka Japan) was added to the cells infected with adenoviruses expressing the WT and CTF TDP-43 at final concentrations of 1–200 μmol/L in the cell viability assay and 50 μmol/L in the sequence analysis assay. Edaravone was prepared in DMSO (100%), and the final concentration of DMSO was set at 0.1% in both vehicle and Edaravone treatments. Twenty-four hours later, the cells received a medium either containing or not containing 20 μmol/L ethacrynic acid (#SML1083; Sigma) and were further incubated for 24 h.
Soejima-Kusunoki A., Okada K., Saito R, & Watabe K. (2022). The Protective Effect of Edaravone on TDP-43 Plus Oxidative Stress-Induced Neurotoxicity in Neuronal Cells: Analysis of Its Neuroprotective Mechanisms Using RNA Sequencing. Pharmaceuticals, 15(7), 842.
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9

Recombinant H-PGDS Enzymatic Assay

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Human recombinant H-PGDS was a kind gift from Professor Bengt Mannervik (Uppsala, Sweden). Ethacrynic acid, cibacron blue, CDNB, GSH were products of Sigma Aldrich. All chemicals unless stated otherwise were purchased from Sigma-Aldrich (Steinheim, Germany).
Moyo R., Chimponda T, & Mukanganyama S. (2014). Inhibition of hematopoietic prostaglandin D2 Synthase (H-PGDS) by an alkaloid extract from Combretum molle. BMC Complementary and Alternative Medicine, 14, 221.
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10

Assay for Oxidative Stress Markers

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1-chloro-2,4-dintrobenzene (CDNB), Anti-DNPH antibody, Reduced glutathione (GSH), Ethacrynic acid, Hydroxynonenal (4-HNE), Glutathione reductase, Dichlorodiphenyltrichloroethane (DDT) and Dichlorodiphenyldichloroethylene (DDE) were purchased from Sigma-Aldrich (USA). All other reagents used were of analytical grades.
Hassan F., Singh K.P., Ali V., Behera S., Shivam P., Das P, & Dinesh D.S. (2019). Detection and functional characterization of sigma class GST in Phlebotomus argentipes and its role in stress tolerance and DDT resistance. Scientific Reports, 9, 19636.
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