Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF

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

• Assay Buffer: 0.1 M MES, pH 6.5
• Recombinant Human Cytosolic beta ‑Glucosidase/GBA3 (rhGBA3) (Catalog # 5969-GH)
• Substrate: 4-methylumbelliferyl-beta -D-glucopyranoside (Sigma, Catalog # M3633), 10 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 rhGBA3 to 1 ng/μL in Assay Buffer.
2. Dilute Substrate to 800 μM in Assay Buffer.
3. Load in a plate 50 μL of 1 ng/μL rhGBA3, and start the reaction by adding 50 μL of 800 μM Substrate. Include a control containing 50 μL of Assay Buffer and 50 μL of 800 μM Substrate.
4. Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), respectively, in kinetic mode for 5 minutes.
5. Calculate specific activity: 

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

• rhGBA3: 0.050 μg
• Substrate: 400 μM