A detailed description of MIP chip preparation procedures can be found elsewhere (Ertürk Bergdahl et al., 2019 (link)). Briefly, α-hemolysin (H9395, Sigma Aldrich) was dissolved in phosphate buffer (10 mM, pH 7.4) to a concentration of 0.1 mg/ml and immobilized onto a pre-treated glass surface, forming a protein (α-hemolysin) mold. A 1.3 µl aliquot of monomer solution [(N-2-Hydroxyethyl methacrylate, 10 %, v/v, Sigma Aldrich, 245801), (Polyethylene glycol dimethacrylate, 50 %, v/v, Sigma Aldrich, 409510), MQ water (40 %, v/v) and (10 mM 1,1’-Azobis cyclohexanecarbonitrile, Sigma Aldrich, 380210)] was placed on the gold surface of a pre-treated Biacore sensor chip (SIA kit Au, Cytiva). Subsequently placed upon the protein mold, all components received 10 min of UV irradiation (Dymax, 400 W, 365 nm). A non-imprinted (NIP) chip was prepared using the same method but excluding the template protein.
1 1 azobis cyclohexanecarbonitrile
Manufactured by Merck Group
Sourced in United States, Spain
1,1'-Azobis cyclohexanecarbonitrile is a chemical compound used in various laboratory applications. It functions as an azo initiator, which is a class of organic compounds that can generate free radicals under specific conditions. The core function of this product is to initiate and facilitate chemical reactions in controlled laboratory environments.
Automatically generated - may contain errors
Lab products found in correlation
N n dimethylformamide (dmf), by Merck Group (2 mentions)
α hemolysin, by Merck Group (1 mentions)
Sia kit au, by Cytiva (1 mentions)
Polyethylene glycol dimethacrylate, by Merck Group (1 mentions)
4 dimethylaminopyridine dmap, by Merck Group (1 mentions)
Xylene, by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Toluene, by Merck Group (1 mentions)
Dimethylformamide, by Merck Group (1 mentions)
Ethylene glycol, by Merck Group (1 mentions)
Methyl methacrylate, by Merck Group (1 mentions)
Butyl methacrylate, by Merck Group (1 mentions)
Hydroxyethyl methacrylate, by Merck Group (1 mentions)
Silica gel, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Isopropanol, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
5 protocols using 1 1 azobis cyclohexanecarbonitrile
1
Molecular Imprinted Polymer Chip Fabrication
2
Synthesis of Functional Methacrylate Polymers
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Dihydro-5-hydroxyl furan-2-one (2H-HBO) was
supplied by Circa group, Australia. Methyl methacrylate (MMA), styrene,
hydroxy ethylmethacrylate (HEMA), and butyl methacrylate (BMA) were
purchased from Sigma-Aldrich and treated with basic alumina to remove
the inhibitor. Methacrylic anhydride (MAN), triethylamine (TEA), and
4-dimethylaminopyridine (DMAP) were purchased from Sigma-Aldrich and
used as received. Azobisisobutyronitrile (AIBN) (provided by CSIRO)
and 1,1′-azobis(cyclohexanecarbonitrile) (purchased from Sigma-Aldrich)
were recrystallized twice from methanol. Benzoyl peroxide (Luperox
A75), dicumyl peroxide, sodium dodecyl sulfate (SDS), and poly(vinyl
alcohol), ammonium persulphate were purchased from Sigma-Aldrich and
used as received. 4-Cyano-4-(((dodecylthio)carbonothioyl)thio)pentanoic
acid (BM1432), chain-transfer agent was supplied by Boron Molecular,
Australia. Solvents such as toluene, ethylene glycol, xylene, dimethylformamide,
and dimethylsulfoxide were purchased from Merck and used as received.
Deionized (Milli-Q, 18 Ω) water was used for emulsion polymerization.
supplied by Circa group, Australia. Methyl methacrylate (MMA), styrene,
hydroxy ethylmethacrylate (HEMA), and butyl methacrylate (BMA) were
purchased from Sigma-Aldrich and treated with basic alumina to remove
the inhibitor. Methacrylic anhydride (MAN), triethylamine (TEA), and
4-dimethylaminopyridine (DMAP) were purchased from Sigma-Aldrich and
used as received. Azobisisobutyronitrile (AIBN) (provided by CSIRO)
and 1,1′-azobis(cyclohexanecarbonitrile) (purchased from Sigma-Aldrich)
were recrystallized twice from methanol. Benzoyl peroxide (Luperox
A75), dicumyl peroxide, sodium dodecyl sulfate (SDS), and poly(vinyl
alcohol), ammonium persulphate were purchased from Sigma-Aldrich and
used as received. 4-Cyano-4-(((dodecylthio)carbonothioyl)thio)pentanoic
acid (BM1432), chain-transfer agent was supplied by Boron Molecular,
Australia. Solvents such as toluene, ethylene glycol, xylene, dimethylformamide,
and dimethylsulfoxide were purchased from Merck and used as received.
Deionized (Milli-Q, 18 Ω) water was used for emulsion polymerization.
3
Synthesis of Peptide-Based Compounds
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The resin Rink amide and the reagents Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Ile-OH, Fmoc-Asn(Trt)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-His(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Cys(Trt)-OH, Fmoc-6-aminohexanoic acid (Fmoc-Ahx-OH), dicyclohexylcarbodiimide, and 1-hydroxy-6-chlorobenzotriazole were purchased from AAPPTec (Louisville, KY, USA). The reagents acetonitrile, trifluoroacetic acid, dichloromethane, diisopropylethylamine, N,N-dimethylformamide, ethanedithiol, isopropanol, methanol, and triisopropylsilane were purchased from Merck (Darmstadt, Germany). SPE columns Supelclean™, 6-maleimidohexanoic acid (6-MhxA), tetrabutylammonium chloride, glycidyl methacrylate (GMA), ethylene dimethacrylate (EDMA), cyclohexanol, dodecanol, 1,1′-azobis(cyclohexanecarbonitrile), silica gel, Tween 20, and 3,3′,5,5′-tetramethylbenzidine (Liquid Substrate System for ELISA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Potassium diacid phosphate, sodium acid phosphate, sodium chloride, and potassium chloride were purchased from AppliChem Panreac (Barcelona, Spain).
4
Synthesis of Functional Polymers
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Styrene (S, ≥99%), sodium azide (NaN3, ≥99%), N,N-dimethylformamide (DMF, ≥99.9%), and 1,1′-azobis(cyclohexanecarbonitrile) (ACHN, ≥98%) were purchased from Sigma-Aldrich (Madrid, Spain). Chloromethyl Styrene (CMS, ≥90%) and sym-dibenzo-1,5-cyclooctadiene-3,7-diyne (DIBOD, > 98%) were supplied by TCI. Tetrahydrofuran (THF, GPC grade) and methanol (MeOH, analytical grade) were purchased from Scharlau (Barcelona, Spain). Benzyl azide (BzA, 94%), S-cyanomethyl-S-dodecyltrithiocarbonate (CDTC, ≥97%), and ethanol (EtOH, 96% v/v) were supplied by Thermo Scientific (Eindhoven, Netherlands), Strem (Bischheim, France), and PanReac AppliChem (Barcelona, Spain), respectively. Unless otherwise specified, all reagents were used as received without further purification. To remove the inhibitor, S and CMS were purified by being passed through a column packed with activated basic alumina.
5
Polymer Synthesis and Characterization
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Methanol, n-hexane, acetone (analytical grade), and triethylamine (TEA) (HPLC grade) were from J.T. Baker (USA). Tetrahydrofuran (THF), deuterated chloroform (CDCl3), methyl methacrylate (MMA, 99%), and dimethyl 2-(aminoethyl) methacrylate (DMAEMA, 98%) were from Sigma-Aldrich (USA) and their polymerization inhibitor, MEHQ, was removed using De-HiBit-200 (Polysciences, Inc., USA) macroreticular ion exchange resin. 2,2′-azobisisobutyronitrile (AIBN, 98%), 1,1′-azobis(cyclohexane carbonitrile) (ACHN, 98%), 2-cyano-2-propyl dodecyl trithiocarbonate (chain transfer agent, CTA), and buffers were from Sigma-Aldrich. NaH2PO4, boric acid, HCl, and NaOH were from Merck (Germany). Isopropyl β-D-1-thiogalactopyranoside (IPTG) and 5-bromo-4-chloro-3-indolyl β-D-galactoside (X-Gal) were from Thermo Scientific (USA). Gel permeation chromatography (GPC) standard for average molar mass determination was Poly(methyl methacrylate) Standard ReadyCal Set, Mp 500-2,700,000, from Sigma Aldrich.
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