Enzyme Assays

Recombinant Mouse Heparan Sulfate 6-O-Sulfotransferase-3/HS6ST3 Activity Assay using Recombinant Mouse HS6ST3 (Catalog # 5406-ST).

Recombinant Mouse Carbohydrate Sulfotransferase 7/CHST7 Activity Assay using Recombinant Mouse CHST7 Protein (Catalog # 5108-ST).

Recombinant Mouse Carbohydrate Sulfotransferase 5/CHST5 Activity Assay using Recombinant Mouse CHST5 Protein (Catalog # 5210-ST).

Recombinant Mouse Carbohydrate Sulfotransferase 1/CHST1 Activity Assay using Recombinant Mouse CHST1 Protein (Catalog # 5355-ST)

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 Meprin alpha Subunit/MEP1A Assay

1. Dilute recombinant human Meprin alpha Subunit (R&D Systems, Catalog # 3220-ZN) to 0.1 mg/mL in TCNB buffer [50 mM Tris, 150 mM NaCl, 10 mM CaCl₂ (Sigma, Catalog # C-5080), 0.05% Brij® 35, pH 7.5] containing 0.1 µg/mL Trypsin. Mix well by gently vortexing.
2. Incubate the mixture at 37° C for 3 hours in a heat block covered with foil. Stop trypsin activity by adding 1 mM AEBSF (R&D Systems, Catalog # EI001).

Recombinant Mouse ACE-2 Assay

1. Dilute recombinant mouse ACE-2 (R&D Systems, Catalog # 3437-ZN) to 0.5 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.