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ÎҲο¼µÄÎÄÏ×£ºRu tris(2,2'-bipyridine-4,4'-dicarboxylic acid) N-hydroxysuccinimide ester
(4). 0.46 g of DCC and 0.238 g of N-hydroxysuccinimide were dissolved
in 3 ml of DMF with stirring and cooled in an ice bath. A solution of 0.38
g of Ru tris(2,2'-bipyridine-4,4'-dicarboxylic acid) (2) was added, and the
mixture was stirred for a few hours. The formed precipitate was removed
by filtration through a syringe filter, and the filtrate containing the active
Ru-complex was used for labeling the substrates. The NHS-ester (3) was
prepared in analogy to (4).
The proteins HSA, IgG, ConA, and Ferritin were obtained from Sigma
Chemical Co. (St. Louis, MO) and used without further purification. The
proteins (10 mg portions) were labeled by adding a 100-fold molar excess
of the Ru-NHS ester in 50 jLI of DMF to 1 ml of stirred protein solution
(0.2 M carbonate buffer, pH 8.3-9.1), followed by a 2-6 h incubation and
purification of the labeled protein by gel filtration chromatography on
Sephadex G-25 or G-50, using 0.1 M PBS, pH 7.2.
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