Recombinant Human MAN2B1 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 Acetate, pH 4.5
• Recombinant Human MAN2B1 (rhMAN2B1) (Catalog # 9826-GH)
• Substrate: 4-Methylumbelliferyl-alpha -D-mannopyranoside (Sigma, Catalog # M3657), 50 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 rhMAN2B1 to 2 ng/μL in Assay Buffer.
2. Dilute Substrate to 400 µM in Assay Buffer.
3. Load 50 μL of 2 ng/μL rhMAN2B1 into the plate, and start the reaction by adding 50 μL of 400 µM Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 400 µM Substrate.
4. Read at excitation and emission wavelengths of 365 nm and 445 nm, respectively, (top read) in kinetic mode for 5 minutes.
5. Calculate specific activity:

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

• rhMAN2B1: 0.1 µg
• Substrate: 200 µM