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3 protocols using baylase rp

1

Enzyme-Catalyzed Peroxidase Assay Protocol

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A commercial peroxidase, Baylase® RP, was kindly donated by Bayer Mexico (Puebla, Mexico). Crystalline silicon was a product from Cemat Silicon (Warsaw, Poland). 10-undecenoic acid, N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide (EDC), N-hydroxysuccinimide (NHS), 2, 2′-azino-bis (3 ethylbenzothiazoline-6-sulphonic acid) (ABTS), Guaiacol, and Remazol brilliant blue were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bradford reagent was from Bio-Rad (Hercules, CA, USA). All other chemical reagents were of analytical grade and were used without further purification.
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2

Enzyme Immobilization on Silicon

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A commercial peroxidase, Baylase® RP, was kindly donated by Bayer Mexico
(Mexico, Federal District, Mexico). Crystalline silicon was a product from Cemat
Silicon (Warsaw, Poland). Glutaraldehyde, 3-aminopropyldiethoxysilane, guaiacol, and
bovine serum albumin were from Sigma-Aldrich (St. Louis, MO, USA). Bradford reagent
was from Bio-Rad (Hercules, CA, USA). All other chemical reagents used in our
experiment were of analytical grade without further purification.
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3

Synthesis and Characterization of Enzyme-Linked Immunosorbent Assay

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Nitric acid (HNO3, 70%), sulphuric acid (H2SO4, >95%), acetic acid (CH3COOH, laboratory reagent grade), ethanol (EtOH, 99%), magnesium powder (99%) hydrochloric acid (HCl, 36%) and sodium hydroxide pellets (NaOH, ≥97.0%) were obtained from Fisher Scientific. 10-Undecenoic acid 98%, 2, 2'-Azino-bis(3ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), 3-aminopropyltriethoxysilane (APTES), N-Hydroxysuccinimide (NHS), N-(3-dimethylamonopropyl)-N' ethylcarbodimiide hydrochloride (EDC), glutaradehyde (GDA, 50%) and peroxidase from horseradish (Type VI-A, salt free, lyophilized powder 950-2000 U/mg) were obtained from Sigma Aldrich. A commercial peroxidase, Baylase® RP; hereafter named Coprinus cinereus peroxidase (CiP), was kindly donated by Bayer Mexico (Puebla, Mexico). Phosphate buffer was prepared by using monobasic potassium phosphate also from Sigma Aldrich. Succinic anhydride was obtained from Fluka. Standard solutions for each element were prepared from 1000 ppm stock solutions purchased from BDH Laboratory Supplies. Distilled-deionized water (ddH2O) with conductivity less than 1 μS cm -1 (25 °C) was used for all experiments.
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