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We describe the use of biotinylated Sepharose, a product of the chromatographic reagent company Sigma, for quantification of protein binding, in our laboratory. For this purpose, we synthesized a high molecular weight agarose matrix (M(r) = 50 kDa) which is conjugated with biotin. Binding experiments of radioactive or nonradioactive compounds to the biotinylated Sepharose are carried out without labeling the compounds and are not subject to false positive results as have been seen with some other reagents. The advantages of using biotinylated Sepharose are: 1) accurate binding of protein; 2) simple; 3) specific; 4) easy to perform; and 5) does not require washing. The technique is highly reproducible and is as efficient as those involving radioactivity. dehors” (w. 80
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