Recombinant Human TACE/ADAM17 Protein, CF

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

• Assay Buffer: 25 mM Tris, 2.5 μM ZnCl₂, 0.005% Brij-35 (w/v), pH 9.0 (note: It is extremely important that the assay solution does not contain salt (CaCl₂, NaCl, Na₂SO₄) because it inhibits TACE activity).
• Recombinant Human TACE/ADAM17 (rhTACE) (Catalog # 930-ADB)
• Substrate: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH₂ (Catalog # ES003) , 2 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 rhTACE to 0.2 ng/µL in Assay Buffer.
2. Dilute Substrate to 20 µM in Assay Buffer.
3. In a plate load 50 µL of 0.2 ng/µL rhTACE and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 µL Substrate.
4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
5. Calculate specific activity:

     *Adjusted for Substrate Blank
     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).