Recombinant Human alpha-Galactosidase A/GLA Protein, CF

• 6 months from date of receipt, -20 to -70 °C as supplied.
• 3 months, -20 to -70 °C under sterile conditions after opening.

• Assay Buffer: 50 mM Sodium Citrate, 50 mM NaCl, pH 4.0
• Recombinant Human alpha ‑Galactosidase A/GLA (rhGLA) (Catalog # 6146-GH)
• Substrate: 4-methylumbelliferyl-alpha -D-galactopyranoside (Sigma, Catalog # M7633), 6.7 mM Stock in DMSO
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
• Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1. Dilute rhGLA to 4.0 ng/µL in Assay Buffer.
2. Dilute Substrate to 800 µM in Assay Buffer.
3. Load into plate 50 µL of 4.0 ng/µL rhGLA, and start the reaction by adding 50 µL of 800 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate.
4. Read at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in kinetic mode for 5 minutes.
5. Calculate the specific activity:

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Methylumbelliferone (Sigma, Catalog # M1381).

• rhGLA: 0.200 µg
• Substrate: 400 µM