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Recombinant Mouse TACE/ADAM17 Protein, CF

Catalog #: 2978-AD
5 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
2978-AD-010
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Recombinant Mouse TACE/ADAM17 Protein, CF Summary

Product Specifications

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.
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.
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
Accession #
AAC62934
N-terminal Sequence
Analysis
Pro18, Asp59 & Arg215
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.
Predicted Molecular Mass
63 kDa, 40 kDa, 23 kDa, 18 kDa
SDS-PAGE
76 kDa, 63 kDa, 21 kDa and 18 kDa, reducing conditions

Product Datasheets

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2978-AD

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

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
Entrez Gene IDs
6868 (Human); 11491 (Mouse)
Alternate Names
ADAM 17; ADAM metallopeptidase domain 17; ADAM metallopeptidase domain 18; ADAM17; ADAM18; CD156b antigen; CD156b; CSVP; disintegrin and metalloproteinase domain-containing protein 17; EC 3.4.24.86; MGC71942; Snake venom-like protease; TACE; TACEcSVP; TNF-alpha convertase; TNF-alpha converting enzyme; TNF-alpha-converting enzyme; tumor necrosis factor, alpha, converting enzyme

Citations for Recombinant Mouse TACE/ADAM17 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Regulation of the Receptor Tyrosine Kinase AXL in Response to Therapy and Its Role in Therapy Resistance in Glioblastoma
    Authors: L Scherschin, M Prem, I Kremenetsk, I Tinhofer, P Vajkoczy, AG Karbe, JS Onken
    International Journal of Molecular Sciences, 2022-01-17;23(2):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. A-Disintegrin and Metalloproteinase (ADAM) 17 Enzymatically Degrades Interferon-gamma
    Sci Rep, 2016-08-30;6(0):32259.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  3. The role of metalloproteinase ADAM17 in regulating ICOS ligand-mediated humoral immune responses.
    Authors: Marczynska J, Ozga A, Wlodarczyk A, Majchrzak-Gorecka M, Kulig P, Banas M, Michalczyk-Wetula D, Majewski P, Hutloff A, Schwarz J, Chalaris A, Scheller J, Rose-John S, Cichy J
    J Immunol, 2014-08-08;193(6):2753-63.
    Applications: Bioassay
  4. Canonical transforming growth factor-beta signaling regulates disintegrin metalloprotease expression in experimental renal fibrosis via miR-29.
    Authors: Ramdas V, McBride M, Denby L, Baker A
    Am J Pathol, 2013-10-06;183(6):1885-96.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  5. Strain-induced differentiation of fetal type II epithelial cells is mediated via the integrin alpha6beta1-ADAM17/tumor necrosis factor-alpha-converting enzyme (TACE) signaling pathway.
    Authors: Wang Y, Huang Z, Nayak P, Matthews B, Warburton D, Shi W, Sanchez-Esteban J
    J Biol Chem, 2013-07-25;288(35):25646-57.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay

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