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13 protocols using ethyl acrylate

1

Synthesis and Spin-Coating of PEA and PMA

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PEA and PMA polymers were synthetized by radical polymerization of ethyl acrylate and methyl acrylate (Sigma, St. Louis, MO), initiated by benzoin at 1 wt% and 0.35 wt%, respectively. Samples were then dried by vacuum extraction to constant weight and solubilized in toluene 2.5% w/v for PEA and 6% w/v for PMA. PEA and PMA solutions were then spin coated onto 12 mm round glass coverslips for 30 sec at a velocity of 2000 and 3000 rpm, respectively, with an acceleration of 3000 rpm/sec. Excess toluene was removed by placing coverslips under the vacuum for 2 h at 60°C.
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2

Synthesis of N-Methyl-4-chloropyridinium Triflate

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Methyl triflate, 2-chloropyridine, cyclohexane-1,2-diamine, pyridine-2,6-diamine, ethane-1,2-diamine, aryl halides, ethyl acrylate and styrene purchased from Sigma Aldrich were used. Solvents were dried using standard procedures. N-Methyl-4-chloropyridinium triflate was synthesized by the reported method.27 (link)
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3

Synthesis of Crosslinked Poly(ethyl acrylate) Films

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A series of poly(ethyl acrylate) polymer films were obtained by radical polymerization. Briefly, ethyl acrylate (99%, Sigma-Aldrich) monomer was mixed with 0.5 wt.% benzoin (98% pure Scharlab) as photoinitiator and different proportions (0, 1, 2, 3, 5 and 10 wt.%) of ethylene glycol dimethacrylate (EGDMA) (98%, Sigma-Aldrich) as crosslinker. The reaction was carried out in ultraviolet light for 24 h. After polymerization, samples were washed with ethanol in a Soxhlet extractor for 24 h in order to remove low molecular weight substances, except the sample with 0% of EGDMA, which was dried in a vacuum to constant weight. The 2D substrates will be identified hereinafter as PEA-X%, in which X is the percentage of EGDMA. The films obtained were approximately 1 mm thick.
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4

Bionanohybrid Synthesis and Characterization

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Butyl-Sepharose® 4 Fast Flow from GE Healthcare (Uppsala, Sweden), 2-dipyridyldisulfide (2-PDS), dithiothreitol (DTT), sodium borohydride, dimethylsulfoxide (DMSO), dimethylformamide (DMF), dimethylaminopyridine (DMAP), diisopropylethylamine (DIPEA), p-nitrophenylpropionate (pNPP), Pd(OAc)2, ethyl-acrylate, iodobenzene, cis and trans ethyl cinnamate, and triethylamine were from Sigma (St. Louis, MO, USA). Sucrose monolaurate was from Mitsubishi-Kagaku (Tokyo, Japan). AcN-Cys(SH)-Phe-Gly-Phe-Gly-Phe-CONH2 was purchased from Isogen (De Meern, Netherlands). Inductively coupled plasma atomic emission spectrometry (ICP-OES) of the acidic digestion of the solid powder of bionanohybrid was performed on a Perkin Elmer OPTIMA 2100 DV equipment. The transmission electron microscopy (TEM), high resolution TEM microscopy (HRTEM) were performed on a JEOL 2100F microscope equipped with an EDX detector INCA x-sight (Oxford Instruments, Tokyo, Japan). The X-ray diffraction (XRD) pattern was obtained using a Texture Analysis Diffractometer D8 Advance (Bruker) with Cu Kα radiation.
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5

Skin Sensitization Assay Chemicals

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1-chloro-2,4-dinitrobenzene (DNCB, 99.8%, CAS 97-00-7), cinnamaldehyde (98.4%, CAS 104-55-2), ethyl acrylate (100%, CAS 140-88-5), glutaraldehyde (25%, CAS 111-30-8), isoeugenol (99%, CAS 97-54-1), methyl salicylate (99.4%, CAS 119-36-8), ammonium hydroxide solution (28–30%), bovine serum albumin (BSA), DL-dithiothreitol (DTT), and deferoxamine mesylate salt were supplied by Sigma-Aldrich Corporation (NSW, Australia), high performance liquid chromatography (HPLC) grade methanol and acetonitrile were supplied by Merck (Darmstadt, Germany), sodium hydroxide and ammonium acetate were supplied by Chem-Supply (SA, Australia). Sodium phosphate dibasic and monosodium phosphate were purchased from ThermoFisher Scientific (VIC, Australia).
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6

Emulsion Copolymerization of Acrylic Monomers

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Acrylic monomers such as methyl methacrylate (MMA, 99%), ethyl acrylate (EA, 99%) and itaconic acid (IA, 99%) were purchased from Sigma-Aldrich, Schnelldorf, Germany and used to synthesize the polymers via emulsion copolymerization. Disponil FES 77, sodium lauryl sulphate SLS 101 and Texapon P, products of BASF (Ludwigshafen, Germany) were used as co-emulsifiers special for the manufacture of finely dispersed and electrolyte–stable emulsions. Sodium bicarbonate (NaHCO3 > 99%, Sigma-Aldrich, Steinheim, Germany), ammonium persulfate (APS, >98%, Sigma-Aldrich, Steinheim, Germany) were used as reaction constituents as buffer agent and initiator, respectively. All chemicals were used as received without any further purification. Distilled water was used to as solvent to perform the reactions. The synthesis experiments were performed in a 4 necked 250 mL glass reactor equipped with a condenser, mechanical mixer, nitrogen inlet and dropping funnel.
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7

Polymer Film Synthesis and Protein Adsorption

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PEA and PMA polymers were synthesized by radical polymerization of ethyl acrylate and methyl acrylate (Sigma, St. Louis, MO), initiated by benzoin at 1 wt% and 0.35 wt%, respectively. PEA and PMA were then dried by vacuum extraction to constant weight and solubilized in toluene 2.5% w/v and 6% w/v respectively. PEA and PMA films (~1μm) were obtained by spin-casting polymer solutions on glass coverslips (12 or 25mm) at 2000 (PEA) and 3000 (PMA) rpm for 30 seconds, with an acceleration of 3000 rpm/sec. Samples were dried in vacuum for 2 h at 60oC to remove excess toluene. Polymer surfaces were coated with either natural mouse laminin 111 (LM; Invitrogen) or human plasma FN (Sigma) solutions, in Dulbecco’s phosphate-buffered saline (DPBS) at concentrations 2, 10, 20, 50, and 100 μg/mL for 1h and then rinsed with DPBS before use. Mouse plasma fibronectins carrying mutations in the FNIII10 module were also used. ΔRGD FN (FN without the RGD sequence) and syn FN (FN with a mutation in the synergy sequence DRVPPSRN>DAVPPSAN) were generated and purified as previously reported [37 , 38 (link)]. The amount of adsorbed protein was calculated via depletion assay using the bicinchoninic acid working reagent (Thermo Fisher Scientific, Waltham, MA) as previously reported [36 (link)].
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8

Synthesis of Zeolite NaA and Organic Compounds

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Jiangsu Nine Heaven High-Tech
provides zeolite NaA (≥99.0%). KNO3 (≥99.0%),
ethyl acrylate (≥98.0%), ethanol (≥99.7%), and Hammett
indicator set are purchased from Sigma-Aldrich. 1-Propanol (≥99.0%)
was purchased from J&K. All chemicals were used without further
purification.
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9

Synthesis and Characterization of Poly(ester-co-acrylic) Elastomers

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Acetone (99%),
deuterated solvents for NMR analysis (chloroform-d 99.8 atom % D, dimethylsulfoxide (DMSO)-d6 99.9 atom % D, mEthanol-d4 99.8 atom
% D, and acetone-d6 99.9 atom % D), dicyclopentadiene,
acetic anhydride (95%), isopropenyl acetate (99%), ethyl acrylate,
and butyl acrylate were purchased from Sigma-Aldrich (St. Louis, USA). dl-LA (90%), EG (technical grade), and triethylamine (99%) were
purchased from Acros (Thermo Fischer Scientific; Waltham, USA). Ethanol
(96%) and diethyl ether (96%) were purchased from Scharlau (Barcelona,
Spain). Hydrochloric acid (38%) was purchased from Baker (Phillipsburg,
USA). Glacial acetic acid (≥99.5%) was purchased from Labkem
(Dublin, Ireland). Novozyme 435 was kindly donated by Novozymes company
(Madrid, Spain). Animal fat was kindly donated by “Subproductos
Cárnicos Echevarria y Asociados S.L” (Cervera, Spain).
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10

Mechanofluorescent Anthracene Maleimide Probe

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Reagents: Ethyl acrylate (EA), 1,4-butanediol diacrylate (BDA) and 2-Ethyl acrylate (EA), 1,4-butanediol diacrylate (BDA) and 2-hydroxy-2-methylpropiophenone (HMP) were purchased from Sigma-Aldrich. EA and BDA were purified via elution on activated alumina, and HMP used as received. The mechanofluorescent πextended anthracene maleimide probe (DACL) was synthesized and purified according to previously established procedures [29] . All reagents were sparged with N 2 for 45 min and transferred to a N 2 -filled glovebox.
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