Zn(NO3)2·6H2O, (98%) and H2BDC, (98%) were bought from Sigma-Aldrich (Burlington, MA, USA), DMF, (≥99.8%) was purchased from Merck (Darmstdt, Germany), (DABCO, 98%) was purchased from Alfa Aesar (Lancashire, UK). All other compounds used throughout this study were of an analytical grade. The electrochemical experiments were performed using a three-electrode system-CHI model 824B workstation with a screen printed carbon electrode (SPCE)/chemically modified SPCE as a working electrode, Ag/AgCl (in 3 M KCl) as a reference electrode, and Pt wire as an auxiliary electrode. SPCE was purchased from Zensor R&D (Taichung, Taiwan). A phosphate buffer solution (0.1 M, pH 7 PBS) was prepared by mixing 0.1 M, NaH2PO4, and Na2HPO4. To compare the various electrodes performance, 5 mM of FeCN solution used as a probe. Briefly, the FeCN solution was prepared using 5 mM of K3[Fe(CN)6] and K4[Fe(CN)6], and 0.1 M, KCl used as a supporting electrolyte. For H2O2 detection, 100 mM of H2O2 prepared from 9.8 M of H2O2 (30 wt%).
Dabco
Manufactured by Thermo Fisher Scientific
Sourced in Germany
DABCO (1,4-Diazabicyclo[2.2.2]octane) is a heterocyclic organic compound commonly used as a laboratory reagent. It functions as a catalyst and facilitates various chemical reactions in research and analytical settings.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
H2bdc, by Merck Group (1 mentions)
Zn no3 2 6h2o, by Merck Group (1 mentions)
N n dimethylformamide (dmf), by Merck Group (1 mentions)
Hplc grade meoh, by Avantor (1 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Mowiol 4 88, by Thermo Fisher Scientific (1 mentions)
Mouse anti ki 67, by Leica (1 mentions)
Neutral red, by Merck Group (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Acridine orange, by Merck Group (1 mentions)
Mowiol 4 88, by Merck Group (1 mentions)
Concanavalin a (cona), by Thermo Fisher Scientific (1 mentions)
Pertex mounting medium, by CellPath (1 mentions)
4 protocols using dabco
1
Electrochemical Characterization of Modified Electrodes
2
Synthesis of Phenylcarbamates via Isocyanate Reactions
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The preparation of methyl phenylcarbamate and butyl phenylcarbamate was performed by reacting phenyl isocyanate (PhNCO, ≥99%, Acros Organics, Geel, Belgium) with the corresponding alcohol, methanol (MeOH, HPLC grade, VWR Chemicals, Debrecen, Hungary) and butan-1-ol (BuOH, ≥99%, VWR Chemicals, Debrecen, Hungary) in excess, respectively, and acetonitrile (ACN, ≥99%, VWR Chemicals, Debrecen, Hungary) was used as a solvent. To achieve a low water content, BuOH (≥99%, VWR Chemicals, Debrecen, Hungary) and ACN (≥99%, VWR Chemicals, Debrecen, Hungary) were stored in over 20% (m/V) activated molecular sieves (3 Å, beads, VWR Chemicals, Debrecen, Hungary) for at least two days [18 (link)]. The products were purified by flash column chromatography (hexane/ethyl acetate) on silica. The studied catalysts were 1,4-diazabicyclo[2.2.2]octane (DABCO, 98%, Alfa Aesar, Kandel, Germany); 1,2-dimethylimidazole (1,2-DMI, 98%, Alfa Aesar, Kandel, Germany); N-ethylmorpholine (NEM, 98%, Alfa Aesar, Kandel, Germany).
3
Quantifying Apoptosis and Proliferation in OCT Biofilms
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OCT embedded biofilm wounds, sectioned at 10 μm, were blocked in appropriate serum and incubated in primary antibodies O/N at 4°C. The antibodies used were the early apoptosis marker, goat anti-caspase 3 (R&D Systems), and the cell-proliferation marker, mouse anti-Ki67 (Novocastra). Alexa Fluor conjugated secondary antibodies (Thermo Fisher Scientific) were used to detect antibody binding. Sections were counterstained in DAPI and mounted in Mowiol 4-88 with DABCO (Thermo Fisher Scientific). Fluorescent images were taken as above using the DAPI, FITC and TEXAS RED filters.
4
Visualizing Porcine Wound Biofilm
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Samples embedded in optimal cutting temperature media (OCT, CellPath) were cryo-sectioned at 10 μm. Gram-Twort, Acridine Orange (AO; Sigma-Aldrich), and Concanavalin A (ConA, Thermo Fisher Scientific) staining were used to visualize porcine wound biofilm load. A modified Gram-Twort stain was carried out. Here, sections were fixed in methanol at -20°C for 10 min, stained with Crystal Violet and Gram’s Iodine solutions (both Sigma-Aldrich), differentiated in 2% (v/v) acetic-alcohol and counterstained with a 0.2% (w/v) neutral red and 0.2% (w/v) fast green (9:1) solution (Sigma-Aldrich). Sections were differentiated again, rapidly dehydrated in 100% ethanol and mounted with Pertex® mounting medium (CellPath). Images were captured at ×100 magnification on a Nikon E400 microscope with SPOT camera (SPOT imaging). For fluorescent visualization, methanol-fixed sections were incubated in AO solution (2 mg/ml) for 5 min at RT and rinsed in dH2O, or incubated in ConA (50 μg/ml) at 4°C O/N and counterstained with DAPI (Thermo Fisher Scientific). Mowiol 4-88 (Sigma-Aldrich) containing DABCO (Thermo Fisher Scientific) was used for mounting. Fluorescent images were taken on a Zeiss Axio Vert. A1 microscope with AxioCam| cm1 camera (Carl Zeiss Microscopy Ltd) at ×40 magnification. Gram-Twort biofilm thickness analysis was performed in ImageJ v.1.6 (NIH).
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