Recombinant Human Monoglyceride Lipase Protein, CF

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

• Assay Buffer: 50 mM Tris, pH 8.0
• Ethanol, Absolute
• Recombinant Human Monoglyceride Lipase (rhMGLL) (Catalog # 7930-MG)
• Substrate:  4-Nitrophenyl butyrate (PNPB) (Sigma, Catalog # N9876), 100 mM stock in Acetone
• 96-well Clear Plate (Costar, Catalog # 92592)
• Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

1. Dilute rhMGLL to 0.8 ng/μL in Assay Buffer.
2. Dilute Substrate to 2 mM using 40% ethanol.
3. Load 50 μL of 0.8 ng/μL rhMGLL into the plate, and start the reaction by adding 50 μL of 2 mM Substrate.  Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 2 mM Substrate.
4. Read at an absorbance of 410 nm in kinetic mode for 5 minutes.
5. Calculate specific activity:

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Nitrophenol (4-NP) (Sigma, Catalog # 241326).

• rhMGLL: 0.04 μg
• PNPB: 1 mM