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Enzyme Assays

Recombinant Human Carbohydrate Sulfotransferase 1/CHST1 Activity Assay

Figure 1. Representative diagram of substrate and product migration.
  1. Dilute recombinant human CHST1 (R&D Systems, Catalog # 5316-ST) to 20 µg/mL, 10 µg/mL, 5 µg/mL, and 2.5 µg/mL in Assay Buffer 1.

Recombinant Human BMP-1/PCP Assay

  1. Dilute recombinant human BMP-1 (R&D Systems, Catalog # 1927-ZN) to 20 µg/mL in BMP-1 assay buffer [25 mM HEPES (Sigma, Catalog # H3375 or Amresco, Catalog # 0511), 0.01% Brij® 35 (Sigma, Catalog # B4184), pH 7.5].
  2. Dilute the substrate, Mca-Y-V-A-D-A-P-K(Dnp)-OH (R&D Systems, Catalog # ES007) to 20 µM with BMP-1 assay buffer.
  3. Load a plate into the plate reader.

Recombinant Human ACE-2 Assay

  1. Dilute recombinant human ACE-2 (R&D Systems, Catalog # 933-ZN) to 0.2 µg/mL in ACE-2 assay buffer [75 mM Tris (Amresco, Catalog # 0826), 1 M NaCl (J.T. Baker, Catalog # 3624-07), pH 7.5].
  2. Dilute the substrate, Mca-Y-V-A-D-A-P-K(Dnp)-OH (R&D Systems, Catalog # ES007) to 20 µM with ACE-2 assay buffer.
  3. Load a plate into the plate reader.
  4. Add 50 µL of enzyme to each well.

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