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Establishment of a detergent-solubilized γ-secretase assay with a notch-based substrate. (A) Cleavage of V1711 with γ-secretase was monitored by Western blot to cleaved NICD. Cleavage was linear with respect to time for the first 45 min. (B) The reaction rate doubled with doubling enzyme concentration (mean ± SD). (C) The amount of active γ-secretase enzyme used in the detergent-soluble assay was titrated with the highly potent inhibitor LY411,575.
