Recombinant Mouse TACE/ADAM17 Protein, CF

R&D Systems | Catalog # 2978-AD

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse TACE/ADAM17 Protein (2978-AD)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

AAC62934

Structure / Form

Recombinant Mouse TACE/ADAM17 is prone to proteolytic cleavage at C-terminus. The poly-His tag may not be present in the preparation.

Applications

Enzyme Activity
View all TACE/ADAM17 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse TACE/ADAM17 protein
Pro18-Asp563 and Arg215-Asp563, both with a C-terminal 10-His tag, Pro18-Arg214 and Asp59-Arg214

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Pro18, Asp59 & Arg215

Predicted Molecular Mass

63 kDa, 40 kDa, 23 kDa, 18 kDa

SDS-PAGE

76 kDa, 63 kDa, 21 kDa and 18 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic peptide substrate Mca-PLAQAV-Dpa-RSSSR-NH2 (Catalog # ES003).
The specific activity is >500 pmol/min/µg, as measured under the described conditions.  Note: It is extremely important that the assay solution does not contain salt as it inhibits TACE activity.

Formulation, Preparation, and Storage

2978-AD
Formulation Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
Reconstitution

Reconstitute at 100 μg/mL in sterile, deionized water.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: TACE/ADAM17

Tumor necrosis factor alpha (TNF-alpha ) converting enzyme (TACE) is a member of the ADAM family that contains a disintegrin and metalloprotease-like domain. The domain structure of mouse TACE consists of pro (aa 18-214), catalytic (aa 215-474), disintegrin-like (aa 475-563), cysteine-rich (aa 564-602), Crambin-like (aa 603-671), transmembrane (aa 672-692), and cytoplasmic (aa 693-827) (1, 2). Recombinant mouse TACE ends after the disintegrin-like domain and consists of several forms as described.  In addition to its ability to release the 17 kDa extracellular form of TNF-alpha from the 26 kDa membrane-anchored TNF-alpha, TACE also plays an essential role in shedding ectodomains from a variety of proteins such as L-selectin, transforming growth factor-alpha, amyloid protein precursor, and Notch1 receptor. TACE mRNA is present in virtually every tissue and TACE protein resides both on the cell surface and in the cell (3).

References

  1. Cerretti, D.P. et al. (1999) Cytokine 11:541.
  2. Mizui, Y. et al. (1999) Gene 233:67.
  3. Black, R.A. (2004) Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. pp. 726, Academic Press, San Diego.

Long Name

TNF-alpha Converting Enzyme

Alternate Names

ADAM17, CD156b

Entrez Gene IDs

6868 (Human); 11491 (Mouse)

Gene Symbol

ADAM17

UniProt

AAC62934

Additional TACE/ADAM17 Products

Product Documents for Recombinant Mouse TACE/ADAM17 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse TACE/ADAM17 Protein, CF

For research use only

Citations for Recombinant Mouse TACE/ADAM17 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse TACE/ADAM17 Protein, CF (2978-AD):

Materials
  • Assay Buffer: 50 mM Tris, pH 9.0
  • Recombinant Mouse TACE/ADAM17 (rmTACE) (Catalog # 2978-AD)
  • 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 rmTACE 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 rmTACE to wells, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank of 50 µL Assay Buffer and 50 µL of 20 µM 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:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

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

Per Well:

  • rmTACE: 0.01 µg
  • Substrate: 10 µM

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