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2342514979
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note
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Field emission Scanning Electron Microscope (FESEM) images were obtained on a JEOL JSM-6332 (JEOL USA, Peabody, MA) instrument at an accelerating voltage of 5 kV.
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12
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2342520874
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note
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-4 Pa. Electron beam bombardment evaporated pure Si (99.9995%) (Alfa Aesar, Ward Hill, MA) onto the substrate at a rate of 5.5 Å/s.
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13
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51249163843
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2342586264
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note
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The silanol functional groups were coupled to APTES with the use of 1% (v/v) solution of APTES in anhydrous toluene at 40°C. After 1 h, excess APTES was removed with toluene (200 ml) and then with a mixture of acetonitrile/water (50%, v/v, 200 ml), and a final wash was completed with excess Milli-Q water (200 ml). The chip was then allowed to dry in air.
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2342618404
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note
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2 projected area on the Si wafer base.
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2342540630
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note
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Polymer coupling to APTES-treated supports was achieved by dipping the supports in a 1% (w/v) PMA solution in anhydrous acetone (20 ml) for 10 min. Excess layers of PMA were removed by dipping each support in pure anhydrous acetone three times. SBP (Sigma Chemical Co., St. Louis, MO) was then coupled to the PMA by incubating 1 mg/ml SBP (0.1 M, pH 7.0 phosphate buffer) for 2 h at room temperature. To remove loosely bound SBP, the supports were washed (3 X 1.5 ml) with the same buffer and then incubated in 1.5 ml buffer for 2 h.
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33947484220
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Y. Levin, M. Pecht, L. Goldstein, and E. Katchalski, Biochemistry 3, 1905 (1964).
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2342485206
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note
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The thickness of the layers deposited on silicon wafers was measured after each treatment with an M-44 Ellipsometer (J. A. Woollam Co., Inc., Lincoln, NE). The reported value is an average of three readings obtained at an incident angle of 75°. The values of the refractive index used for APTES, PMA, and SBP were 1.45, 1.51, and 1.51, respectively.
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2342580335
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2 (250 μl, 0.1 M sodium phosphate buffer, pH 7, with 20% (v/v) dimethyl formamide), and the fluorescence was monitored as described above.
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2342582332
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2,2-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) is a sensitive substrate for peroxidase activity and is useful for measuring the activity of sub-ng/ml concentrations of SBP. The initial rate of SBP-catalyzed oxidation of ABTS was measured by monitoring the increase in the absorbance of the end product at 405 nm with a bioassay reader.
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27
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0000832894
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edited by L. B. Wingard, Jr., E. Katchalski-Katzir, and L. Goldstein, Academic Press, New York
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