Recombinant Human TRACP/PAP/ACP5 Protein, CF

• 3 months from date of receipt, 2 to 8 °C as supplied.
• 1 month, 2 to 8 °C under sterile conditions after opening.

• Assay Buffer: 50 mM NaOAc, pH 5.0
• Recombinant Human TRACP/PAP/ACP5 (rhACP5) (Catalog # 3948-AP)
• Substrate: p-Nitrophenyl phosphate (Sigma-Aldrich, Catalog # N2765)
• 96-well Clear Plate (Costar, Catalog # 92592)
• Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
• NaOH, 0.2 M in deionized water

1. Dilute rhACP5 to 0.1 μg/mL in Assay Buffer.
2. Dilute Substrate to 2 mM in Assay Buffer.
3. In a plate combine 50 μL of rhACP5 and 50 μL of 2 mM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL of 2 mM Substrate.
4. Incubate reaction at room temperature for 10 minutes in the dark.
5. Add 100 μL of 0.2 M NaOH to stop the reaction and develop the color.
6. Read (top read) absorbance in endpoint mode at 410 nm.
7. Calculate specific activity:

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

• rhACP5: 0.005 μg
• Substrate: 0.5 mM