Mouse recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4), recombinant murine FLT3-Ligand (FLT3L) and HGF were obtained from Peprotech (Rocky Hill, NJ, USA). FM™ 4-64 Dye and Alexa Fluor 488-conjugated ovalbumin (OVA) was procured from Molecular Probes (Eugene, OR, USA). Calphostin C, superoxide dismutase (SOD), diphenyleneiodonium chloride (DPI), LY294002, 5-(N-ethyl-N-isopropyl) amiloride (EIPA), phorbol 12-myristate 13-acetate (PMA), cytochalasin D, lipopolysaccharide (LPS), and OVA were purchased from Sigma-Aldrich (St. Louis, MO, USA). Protease and phosphatase inhibitor cocktail tablets were purchased from Roche Diagnostics GmbH (Mannheim, Germany). FITC-OVA and dihydroethidium (DHE) were obtained from Thermofisher Scientific (Grand Island, New York, NY, USA). EUK-134 was purchased from Cayman Chemical (Ann Arbor, MI, USA). L-012 was obtained from Wako Chemicals (Richmond, VA, USA). PE/Cy7-CD11c (clone N418), APC-CD11c (clone N418), FITC-MHCII (clone M5/114.15.2), APC-CD86 (clone GL1) monoclonal antibodies, and respective isotype controls were purchased from Thermofisher Scientific.
Euk 134
Manufactured by Cayman Chemical
Sourced in United States
EUK-134 is a synthetic antioxidant compound. It functions as a mimic of the antioxidant enzymes superoxide dismutase and catalase.
Automatically generated - may contain errors
Lab products found in correlation
Mitotempo, by Merck Group (2 mentions)
N acetyl cysteine (nac), by Merck Group (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Protease and phosphatase inhibitor cocktail tablet, by Roche (1 mentions)
Diphenyleneiodonium chloride, by Merck Group (1 mentions)
Fm4 64 dye, by Thermo Fisher Scientific (1 mentions)
Calphostin c, by Merck Group (1 mentions)
Superoxide dismutase, by Merck Group (1 mentions)
Fitc ova, by Thermo Fisher Scientific (1 mentions)
Dihydroethidium dhe, by Thermo Fisher Scientific (1 mentions)
L 012, by Fujifilm (1 mentions)
Cytochalasin d, by Merck Group (1 mentions)
Hepatocyte growth factor (hgf), by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharide, by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
Ab129068, by Abcam (1 mentions)
Thapsigargin, by Thermo Fisher Scientific (1 mentions)
Anti ece1, by Thermo Fisher Scientific (1 mentions)
Hydrogen peroxide, by Thermo Fisher Scientific (1 mentions)
21 protocols using euk 134
1
Murine Dendritic Cell Activation Assay
2
Proteomic Analysis of ET Signaling
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Rat monoclonal anti-pan-ET (GeneTex, Irvine, CA; GTX40860), rabbit polyclonal anti-ECE1 (Life Technologies, Grand Island, NY; PA5-81948), rabbit monoclonal anti-catalase (Abcam, Cambridge, MA; ab209211), rabbit monoclonal anti-NOX2 (Abcam; ab129068), rabbit monoclonal anti-NOX4 (Abcam; ab133303), rabbit polyclonal anti-TRPC3 (Alomone, Jerusalem Israel; ACC-016), rabbit polyclonal anti-ET receptor A (Alomone; AER-001), rabbit polyclonal anti-ET receptor B (Bioss Antibodies, Woburn, MA; bs-4198R), and mouse monoclonal anti-beta actin (Life Technologies; MA515739) were used. Secondary antibodies for Western blot (anti-mouse: 1:5000; anti-rabbit: 1:15000; Abcam) and immunofluorescence (CF488 Donkey anti-mouse and CF555 Donkey anti-Rabbit at 1:400 dilutions; Biotium, Inc, Fremont, CA) were used. Primary antibodies for Western blot and immunofluorescence were at 1:300 and 1:100 dilutions, respectively.
Unless specified, all reagents were purchased from MilliporeSigma (Burlington, MA). EUK 134 (EUK), ionomycin and bosentan (Cayman Chemical; Ann Arbor, MI), CGS 35066 (CGS), and SAR 7334 (SAR) (Tocris; Bristol, UK), apocynin, and thapsigargin (Life Technologies) were used.
Unless specified, all reagents were purchased from MilliporeSigma (Burlington, MA). EUK 134 (EUK), ionomycin and bosentan (Cayman Chemical; Ann Arbor, MI), CGS 35066 (CGS), and SAR 7334 (SAR) (Tocris; Bristol, UK), apocynin, and thapsigargin (Life Technologies) were used.
3
Isolation and Culture of Raw 264.7 Macrophages
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The raw 264.7 macrophage cells (ATCC, Manassas, VA, USA) were maintained in DMEM (high glucose) media containing 10% FBS and 100 U/mL penicillin/streptomycin in a 37 °C incubator supplemented with 5% CO2. To obtain the conditioned media, 3 × 106 cells per 10 cm dish were grown in media used for acinar cell 3D collagen explant culture (Waymouth’s media plus supplements, see below) and supernatants were collected. The conditioned media was freshly prepared for each experiment. The PKD1 antibody (AP06569PU-N) was from Acris (San Diego, CA, USA), the β-actin (A5441) antibody was from Sigma-Aldrich (St. Louis, MO, USA). The secondary HRP-linked anti-mouse or anti-rabbit antibodies were from Jackson ImmunoResearch (West Grove, PA, USA). The recombinant murine CCL5/RANTES and TNFα were purchased from PeproTech (Rocky Hill, NJ, USA). The recombinant TGFα was from R&D Systems (Minneapolis, MN, USA). The hydrogen peroxide was from Thermo Fisher (Rochester, MN, USA) and the EUK134 was from Cayman Chemical (Ann Arbor, MI, USA). The PKD-specific inhibitor kb-NB-142-70 was described previously [14 (link),20 (link)]. The collagenase I was from Millipore/Sigma (St. Louis, MO, USA). The rat tail collagen I was from BD Biosciences (San Diego, CA, USA).
4
Pharmacological Modulation of Cellular Pathways
5
Immunofluorescence Staining of Neural Markers
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Primary antibodies used were: mouse anti-BrdU (G3G4; DSHB), mouse anti-GFAP (Thermo Fisher Scientific), rhodamine-conjugated mouse anti-BrdU (Millipore), mouse anti-nestin (Rat-401), mouse anti-NeuN (Millipore), mouse anti–Nqo-1 (Santa Cruz Biotechnology), mouse anti-p62 (Abcam), mouse anti ubiquitin (Santa Cruz Biotechnology), mouse antivinculin (Sigma-Aldrich), rabbit anti-Atg5 (Novus Biologicals), rabbit anti-Atg16L1 (Abgent), rabbit anti-GFAP (Dako), rabbit anti-Ki67 (Spring Bioscience), rabbit anti-LC3 (Cell Signaling), rabbit anti-NBR1 (Cell Signaling), rabbit anti-Nrf2 (Abcam), rabbit anti-p62 (Enzo), rabbit anti-Sox2 (Millipore), rabbit anti-TAX1BP1 (Abgent), rabbit anti-Fip200 (ProteinTech), rabbit anti-olig2 (Millipore), rat anti-Ki67 (BioLegend), and guinea pig antidoublecortin (anti-DCX; EMD Millipore). Secondary antibodies were goat anti–rabbit IgG-FITC, goat anti–rabbit IgG–Texas red, goat anti–mouse IgG-FITC, goat anti–mouse IgG–Texas red, goat anti–mouse IgG-HRP, and goat anti–rabbit IgG-HRP (Jackson Immunology).
DHE and EUK-8 were purchased from Sigma-Aldrich. EUK-134 was purchased from Cayman Chemical. DCFDA, MitoTracker Red, and MitoTracker Green were purchased from Invitrogen.
DHE and EUK-8 were purchased from Sigma-Aldrich. EUK-134 was purchased from Cayman Chemical. DCFDA, MitoTracker Red, and MitoTracker Green were purchased from Invitrogen.
6
Investigating Cell Signaling Modulators
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Reagents and salts for buffer preparation were purchased from Sigma-Aldrich (MO, USA). Lipofectamine® 2000, media culture and supplements were acquired from Life Technologies (NY, USA). EUK-134 was purchased from Cayman Chemical Company (MI, USA). PX-12, nigericin, and valinomycin were acquired from Tocris (MO, USA). BCECF-AM was acquired from Thermo Fisher Scientific (MA, USA). The 30% hydrogen peroxide solution used in this work was acquired from Merck (Darmstadt, Germany).
7
Metabolomic Analysis of Cellular Responses
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Bafilomycin A1, epoxomicin, bestatin, and EUK134 (chloro[[2,2'-[1,2-ethanediylbis[(nitrilo-κN)methylidyne]]bis[6-methoxyphenolato-κO]]]-manganese) were from Cayman Chemical Company (Ann Arbor, MI, USA). MnTMPyP (Mn(III)tetrakis(1-methyl-4-pyridyl)porphyrin) was from ENZO life sciences (Farmingdale, NY, USA). 3-Isobutyl-1-methylxanthine (IBMX), dexamethasone (DEX), insulin, hydrogen peroxide, epoxomicin, batimastat, and H218O were from Sigma (St. Louis, MO, USA). Chemical standards used for metabolite identification were from TCI America (Portland, OR, USA) and when unavailable, Sigma.
8
Oxidative Stress Inhibitors Protocol
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The NOX1 inhibitor 4-(2-Aminoethyl)benzenesulfonyl fluoride (AEBSF), the singlet oxygen (1O2) scavenger histidine, the NOS inhibitor N-omega-nitro-L-arginine methylester hydrochloride (L-NAME) and the HOCl scavenger taurine were obtained from Sigma-Aldrich (Schnelldorf, Germany).
The peroxynitrite decomposition catalyst 5-, 10-, 15-, 20-Tetrakis(4-sulfonatophenyl)porphyrinato iron(III) chloride (FeTPPS), was obtained from Calbiochem (Merck Biosciences GmbH, Schwalbach/Ts, Germany).
The catalase mimetic EUK-134 [chloro([2,2′-[1,2-ethanediylbis[(nitrilo-κN)methylidyne]]bis[6-methoxyphenolato-κO]]]-manganese was a product of Cayman (Ann Arbor, Michigan, U.S.A.) and was obtained from Biomol (Hamburg, Germany).
Detailed information on inhibitors has been previously published74 (link),82 (link)–86 (link). The site of action of inhibitors and scavengers has been presented in detail in the Supplementary Material of references #81 and #85.
The peroxynitrite decomposition catalyst 5-, 10-, 15-, 20-Tetrakis(4-sulfonatophenyl)porphyrinato iron(III) chloride (FeTPPS), was obtained from Calbiochem (Merck Biosciences GmbH, Schwalbach/Ts, Germany).
The catalase mimetic EUK-134 [chloro([2,2′-[1,2-ethanediylbis[(nitrilo-κN)methylidyne]]bis[6-methoxyphenolato-κO]]]-manganese was a product of Cayman (Ann Arbor, Michigan, U.S.A.) and was obtained from Biomol (Hamburg, Germany).
Detailed information on inhibitors has been previously published74 (link),82 (link)–86 (link). The site of action of inhibitors and scavengers has been presented in detail in the Supplementary Material of references #81 and #85.
9
Hippocampal Neuron Treatment with Aβ and Modulators
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We prepared hippocampal neurons from day 1 non-Tg as described previously13 (link), culturing neurons in neurobasal medium supplemented with 1× B27, 600 μM l -glutamine, and penicillin–streptomycin. At DIV14, neurons from both Tg mice were treated with 50 nM Aβ in neurobasal medium supplemented with 0.5× B27 for an additional 24 h, with or without EUK-134 (500 nM, Cayman Chemical), SB203580 (1 µM, EMD Chemicals, Inc.), or MitoTEMPO (1 µM, Sigma) pretreatment for 1 h before the addition of Aβ. Vehicle was used as a control in neurobasal medium supplemented with 0.5× B27 for 24 h.
10
Amphibole Asbestos Fiber Characterization
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