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33645839490
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note
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Polymer: 120 mL of 1.1 mM HAuCLj in methanol is mixed with an aqueous solution of GSH (80 mL, 5.0 mM), pH of ~2, resulting in a change from clear yellow to cloudy white, indicating the formation of the polymer; suspension is stirred for an additional 15 min. Clusters: a large excess of NaBHj in water (10 mL, 0.6 M) is added to the stirring suspension, causing a rapid color change to opaque brown/black, and the solution is stirred for an additional 30 min. The resulting solution is evaporated to near dryness, and the cluster compounds are precipitated with Nfldimethylformamide or dilute methanol to remove the diborane and excess GSH/GSSG. Removal of starting materials and byproducts is determined by NMR. The polymer conversion to clusters was quantitative, i.e., no insoluble (bulk) Au or unreacted polymer.
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14
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0021442947
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33645866382
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note
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Electrophoresis: The cleansed product is redissolved ~4 mg/mL in water with 5% glycerol and purified by PAGE using a Bio-Rad PROTEAN II system. Approximately 30 J.L of the sample was loaded onto a 20 cm x 1 mm acrylamide/bis gel (24% T; 7% C) and eluted with 5 mM Tris, 39 mM glycine, pH 8.8 at 150 V for 6 h. The bands formed by electrophoresis were cut out and placed into distilled water. The resulting solution was evaporated to near dryness, an equal volume amount of 2% acetic acid was added, and then precipitation was induced by methanol. This was repeated twice to form a fine black powder. Yield of band 3 (most abundant) was ~40% (by weight) of all clusters made.
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16
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33645878172
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note
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MALDI-MS: The acetate salt of each separated cluster compound was dissolved at a concentration of 3 mg/mL in water and then mixed (1:1, v:v) with a 0.07 M solution of 2,5-dihydroxybenzoic acid (DHB). Four microliters of that solution was vacuum-dried on a polished stainless steel tip, and the 3rd harmonic of a Nd:YAG (355 nm) was used for desorption/ ionization into a reflectron time-of-flight mass spectrometer. ESI-MS: The acetate salt of the most abundant species was dissolved in watermethanol (60:40) to a concentration of 0.24 mg/mL and sprayed at a flow rate of 3 J./L/min into a Sciex API 165 LC/MS quadrupole mass spectrometer.
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33645874903
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