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High-throughput screening requires simple assays that give reliable quantitative results. A microplate assay was developed for reducing sugar analysis that uses a 2,2′-bicinchoninic-based protein reagent. Endo-1,4-β-d-xylanase activity against oat spelt xylan was detected at activities of 0.002 to 0.011 IU ml−1. The assay is linear for sugar concentrations from 0 to 86 μg ml−1 and can also be used to assay protein concentrations (0 to 143 μg ml−1) on the same plate. A variety of temperatures and pH conditions can be used and, after incubation, the assay requires only one detection reagent and one heating step.