Mu αidoa 2s

IDS enzyme activity was measured in a two‐step protocol using the fluorescent substrate MU‐αIdoA‐2S (Carbosynth) and Aldurazyme (Genzyme) as previously described (Lu et al, 2010). Starting material was standardized to 20 μg of total protein or plasma, 40 μg for liver, heart, lung, spleen, and bone marrow, and 60 μg for brain using a BCA assay (ThermoFisher). For β‐hexosaminidase activity, 1 μg of total protein from brain, or 2 μg from spleen and plasma were added to 0.5 mM 4‐methylumbelliferyl‐N‐acetyl‐β‐d‐glucosaminide substrate (Sigma), incubated for 40 min at 37°C, and stopped with 200 μl of 0.2 M carbonate buffer. Fluorescence was measured using the BioTek Synergy HT plate reader (excitation: 360 nm; emission: 460 nm).

IDS enzyme activity was measured in a two-step protocol using the fluorescent substrate MU-αIdoA-2S (Carbosynth) and Aldurazyme (Genzyme), as previously described.⁴⁴ (link) Starting material was standardized to 20 μg total protein or plasma; 40 μg for liver, heart, lung, spleen, and BM; and 60 μg for brain using a BCA assay (Thermo Fisher Scientific). For β-hexosaminidase activity, 1 μg total protein from brain or 2 μg from spleen and plasma were added to 0.5 mM 4-methylumbelliferyl-N-acetyl-β-d-glucosaminide substrate (Sigma-Aldrich), incubated for 40 min at 37°C, and stopped with 200 μL of 0.2 M carbonate buffer. Fluorescence was measured using the BioTek Synergy HT plate reader (excitation 360 nm, emission 460 nm).