Tetraethyl orthosilicate (TEOS, 98.0%), ethanol (C2H5OH), triethyl phosphate (TEP) and hydrochloric acid (HCl) were purchased from Group Chemical Reagent Co., Ltd. (Shanghai, China). Calcium nitrate tetrahydrate (Ca(NO3)2·4H2O, 98%) and manganese nitrate tetrahydrate (Mn(NO3)2·4H2, 98%) were obtained from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Pluronic P123 (Mn∼5,800), poly(D,L-lactide) (PDLLA, 98.0%) and dimethyloxalyl glycine (DMOG, ≥98%) were purchased from Sigma-Aldrich (United States).
Dimethyloxalylglycine
Manufactured by Merck Group
Sourced in United States
Dimethyloxalylglycine is a chemical compound used as a laboratory reagent. It is a cell-permeable inhibitor of the HIF-prolyl hydroxylase enzyme, which plays a role in the regulation of hypoxia-inducible factor (HIF) activity. This compound can be used in research applications where the modulation of HIF signaling is of interest.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Pluronic p123, by Merck Group (1 mentions)
Recombinant murine rankl, by Thermo Fisher Scientific (1 mentions)
Phospho iκbα, by Cell Signaling Technology (1 mentions)
M csf, by Thermo Fisher Scientific (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Raw264.7 cell line, by American Type Culture Collection (1 mentions)
Pyrrolidine dithiocarbamic acid, by Merck Group (1 mentions)
Mouse anti human hif 1α monoclonal antibody, by BD (1 mentions)
Bay11 7082, by Merck Group (1 mentions)
Rabbit anti human β actin monoclonal antibody, by Cell Signaling Technology (1 mentions)
Phenyl n tert butylnitrone, by Merck Group (1 mentions)
Mg132, by Merck Group (1 mentions)
Cobalt 2 chloride cocl2, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Nu7026, by Merck Group (1 mentions)
Ve 821, by Merck Group (1 mentions)
Ku55933, by Merck Group (1 mentions)
Azd7762, by Merck Group (1 mentions)
10 protocols using dimethyloxalylglycine
1
Synthesis of Calcium-Manganese Silicate Composite
2
Modulation of Osteoclastogenesis by Alpha-Ketoglutarate Derivatives
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Alpha-ketoglutaric acid (AKG) disodium salt, dimethyl alpha-ketoglutarate (DM-AKG), and dimethyl oxalylglycine (DMOG) were purchased from Sigma-Aldrich. Recombinant murine RANKL and M-CSF were purchased from Peprotech (Rocky Hill, NJ, USA). Antibodies for phospho-IKKα/β, IKKβ, phospho-IκBα, IκBα, NF-κB (P65), histone H3, and β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies for PHD1, PHD2, and PHD3 were purchased from Abcam (Billerica, MA, USA). The RAW264.7 cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Dulbecco’s Modified Eagle’s Medium (DMEM), α-minimum essential medium (α-MEM), fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Gibco (Rockville, MD, USA).
3
Proteasome Inhibitor BSc2118 Modulates HIF-1α Signaling
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BSc2118, a proteasome inhibitor, was a gift from Dr. Kuckelkorn (Charité Universitätsmedizin). LDL and recombinant human tumor necrosis factor-α (TNFα) were purchased from Invitrogen (Carlsbad, CA) and Peprotech (Rocky Jill, NJ), respectively. The free radical scavenger phenyl-N-tert-butyLnitrone (PBN), the NF-κB inhibitor pyrrolidine dithiocarbamic acid (PDTC), the IκB kinase (IKK) inhibitor Bay 11-7082, the prolyl hydroxylase inhibitor dimethyloxalylglycine (DMOG), and the proteasome inhibitor MG132, were obtained from Sigma-Aldrich (St. Louis, MO). The antibodies used in this study include rabbit anti-human laminin A polyclonal antibody (Sigma-Aldrich); mouse anti-Human HIF-1α monoclonal antibody (BD Transduction Laboratories, San Jose, CA); human/rat HIF-2α antibody (R&D Systems, Minneapolis, MN); rabbit anti-human HIF-1β/ARNT polyclonal antibody, rabbit anti-human NF-κB p65 monoclonal antibody, rabbit anti-human β-actin monoclonal antibody (Cell Signaling Technology, Danver, MA); and anti-HIF-1α (hydroxylated Pro402 and Pro564) antibody (EMD Millipore, Billerca, MA). The NE-PER Nuclear Protein Extraction Reagent Kit was purchased from Pierce (Life Technologies, Frederick, MD).
4
Rat H9c2 Cardiomyocyte Hypoxia Modeling
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Rat (Rattus norvegicus) heart/myocardium cells (H9c2; #CRL-1446, ATCC, Manassas, VA, United States) were cultured and maintained following standard provider protocol. H9c2 cells (1 × 106 cells/ml) were cultured in complete cell culture media [ATCC-formulated Dulbecco’s Modified Eagle’s Medium (DMEM, # 30-2002) supplemented with 10% fetal bovine serum (Sigma, United States)] and were incubated in a humidified atmosphere with 5% CO2 at 37°C, and allowed to grow up to 70–80% confluence before exposure to hypoxia or normoxia. When present, 200 μM (19 (link)–21 (link)) of Cobalt (II) chloride [(CoCl2), # C8661, Sigma, United States] a Hypoxia-inducible transcription factor 1 α (HIF-1 α) inducer, 1 mM of Dimethyloxalylglycine [(DMOG); Sigma, United States, # 400091] a HIF-Hydroxylase Inhibitor, 100 μM (22 (link)) of L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate [(A2P), #8960, Sigma, United States] an essential supplement and reactive oxygen species suppressant (23 (link)) and 1 nM (22 (link), 24 (link), 25 (link)) of Echinomycin (ECM, #SML0477, Sigma, United States) an antitumor antibiotic and potent hypoxia-inducible factor 1α (HIF-1α) the inhibitor was directly added to the culture medium.
5
Kinase Inhibition Assay in Zygotes
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For kinase inhibitor experiments zygotes were incubated after scoring for visible pronuclei at 20 hr post-hCG (corresponding to early/mid G1 phase) in continuous presence of the respective inhibitor [10 μM KU-55933 (Hickson et al., 2004 (link)), Sigma-Aldrich; 10 μM VE-821 (Charrier et al., 2011 (link), Reaper et al., 2011 (link)), Sigma-Aldrich; 10 μM NU7026 (Veuger et al., 2003 (link)), Sigma-Aldrich; 2 mM caffeine (Sarkaria et al., 1999 (link)), Sigma-Aldrich; 100 nM AZD7762 (Zabludoff et al., 2008 (link)), Sigma-Aldrich; 5 μM NSC109555 (Jobson et al., 2007 (link)), Sigma-Aldrich; 10 nM PF-477736 (Blasina et al., 2008 (link)), Sigma-Aldrich]. Incubation in Tet3 inhibitor [1 mM Dimethyloxalylglycine (DMOG) (Amouroux et al., 2016 (link)), Sigma-Aldrich] directly followed zygote isolation at 17-18h post-hCG.
6
Stem Cell Differentiation Protocol
7
Hypoxia Responses in Endothelial Cells
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Human umbilical vein endothelial cells (HUVEC) from pools of donors were purchased from Lonza (Lonza, C2519A) and grown in Endothelial Medium Bullet Kit (EGM-2, Lonza, CC-3162). For viral production, HEK 293T were maintained in Dulbecco's modified Eagle's medium (Gibco, 41966052) supplemented with 50 U/mL penicillin (Gibco, 15140122), 50 µg/mL streptomycin (Gibco, 15140122), 2 mM glutamine (Gibco, 25030123), and 10% (v/v) fetal bovine serum (Gibco, 10270106). All cells were grown at 37°C and 5% CO2 in a humidified incubator and tested regularly for mycoplasma contamination. For hypoxia treatment, cells were grown at 37°C in a 1% O2, 5% CO2, 94% N2 gas mixture in a Whitley Hypoxystation H35 (Don Withley Scientific). Alternatively, hypoxic conditions were mimicked using 500 µM Dimethyloxalylglycine (DMOG, Sigma, D3695). Actinomycin, cycloheximide, puromycin and rapamycin purchased from Sigma, were used at 5 µg/mL, 10 µg/mL, 2 µg/mL, and 20 nM, respectively.
8
Comprehensive Cellular Metabolic Assays
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2-deoxy-D-glucose (2-DG), sodium butyrate (NaBt), sodium valproate (NaVPA), bovine serum albumin (BSA), phenazine methosulfate (PMS), sulforhodamine B (SRB), trichloroacetic acid (TCA), acetic acid, dimethyl sulfoxide (DMSO), cycloheximide (CHX), chloroquine (CQ), dimethyloxalylglycine (DMOG), rhodamine (Rho), 5-bromodeoxyuridine (BrdU) Cell Proliferation Assay Kit, and Lactate Assay Kit were purchased from Sigma-Aldrich, Saint Louis, MO, USA. 3,6-di-O-acetyl-2-deoxy-D-glucose (WP1122) was produced at the MD Anderson Cancer Institute by Professor Waldemar Priebe’s team and was kindly provided for research by the Dermin company, who, based on an exclusive license, conduct developmental studies of the molecule.
3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, inner salt (MTS), was purchased from Promega (Madison, WI, USA). Muse™ Annexin V & Dead Cell Kit was purchased from Calbiochem, Merck Millipore (Darmstadt, Germany). HDAC Activity Colorimetric Assay Kit was purchased from BioVision (Milpitas, CA, USA). All the other reagents and solvents used in this study were of the highest analytical reagent grade.
3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, inner salt (MTS), was purchased from Promega (Madison, WI, USA). Muse™ Annexin V & Dead Cell Kit was purchased from Calbiochem, Merck Millipore (Darmstadt, Germany). HDAC Activity Colorimetric Assay Kit was purchased from BioVision (Milpitas, CA, USA). All the other reagents and solvents used in this study were of the highest analytical reagent grade.
9
Molecular Signaling Mechanisms in Angiogenesis
10
Stimulation and Staining Protocol
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Cells were stimulated with various doses of lipid A Salmonella Minnesota Re595 (VWR), 100ng/ml recombinant mouse interferon-γ (Life Technologies), 0.5mM dimethyloxalylglycine (Sigma-Aldrich) or 10μM Trichostatin A (Sigma-Aldrich). Slide mounting and nuclei staining was performed using Vectashield mounting medium with DAPI (Vector Laboratories).
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