Recombinant Mouse ADAM10 Protein, CF

R&D Systems | Catalog # 946-AD

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Mouse ADAM10 Protein (946-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

Sf 21 (baculovirus)

Accession Number

O35598

Structure / Form

Recombinant Mouse ADAM10 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.

Applications

Enzyme Activity
View all ADAM10 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse ADAM10 protein
Thr215-Glu673, with a C-terminal 10-His tag

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

Thr215

Predicted Molecular Mass

52 kDa

SDS-PAGE

60 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 >5 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

946-AD
Formulation Lyophilized from a 0.2 μm filtered solution in MES, NaCl, CaCl2, ZnCl2, and Brij-35.
Reconstitution Reconstitute at 100 μg/mL in sterile 25 mM Tris, pH 7.5.
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: ADAM10

ADAM10 (also known as Kuzbanian, mammalian disintegrin metalloprotease, myelin-associated metalloproteinase) is a member of the ADAM family that contains a disintegrin and metalloprotease-like domain (1, 2). Like other membrane-anchored ADAMs, ADAM10 consists of the following domains, pro with a cysteine switch and furin cleavage sequence, catalytic with the zinc-binding site and Met-turn expected for reprolysins, disintegrin-like, cysteine-rich, EGF-like, transmembrane, and cytoplasmic. ADAM10 is highly conserved, with 97% amino acid identity between mouse, rat, bovine and human and 45% identity between mouse and Drosophila. The active enzyme processes notch, notch ligand delta, and amyloid protein precursor at the alpha site, playing an important role in neurogenesis (3, 4). It also processes the 26 kDa membrane-anchored pro-tumor necrosis factor-alpha (TNF-alpha ) to the 17 kDa mature TNF-alpha (5). It cleaves myelin basic protein and type IV collagen (6, 7). ADAM10 is widely expressed in tissues and resides both on the cell surface and in the cell (8, 9).

References

  1. Rooke, J. et al. (1996) Science 273:1227.
  2. Pan, D. and Rubin, G.M. (1997) Cell 90:271.
  3. Qi, H. et al. (1999) Science 283:91.
  4. Lammich, S. et al. (1999) Proc. Natl. Acad. Sci. USA 96:3922.
  5. Rosendahl, M.S. et al. (1997) J. Biol. Chem. 272:24588.
  6. Chantry, A. et al. (1989) J. Biol. Chem. 264:21603.
  7. Millichip, M.I. et al. (1998) Biochem. Biophys. Res. Comm. 245:594.
  8. Chantry, A. and Glynn, P. (1990) Biochem. J. 268:245.
  9. Fahrenholz, F.S. et al. (2000) Ann. N.Y. Acad. Sci. 920:215.

Long Name

A Disintegrin and Metalloprotease-like Domain 10

Alternate Names

CD156c, Kuzbanian, MADM

Entrez Gene IDs

102 (Human); 11487 (Mouse)

Gene Symbol

ADAM10

UniProt

O35598

Additional ADAM10 Products

Product Documents for Recombinant Mouse ADAM10 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 ADAM10 Protein, CF

For research use only

Citations for Recombinant Mouse ADAM10 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse ADAM10 Protein, CF (946-AD):

Materials
  • Assay Buffer: 25 mM Tris, 2.5 µM ZnCl2, 0.005% (w/v) Brij-35, pH 9.0
  • Recombinant Mouse ADAM10 (rmADAM10) (Catalog # 946-AD)
  • Substrate: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH2 (Catalog # ES003)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmADAM10 to 2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. In a plate load 50 µL of 2 ng/µL rmADAM10, and start the reaction by adding 50 µL of 20 µM Substrate to wells. Include a Substrate Blank containing 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 30 minutes at 37 °C.
  5. Calculate specific activity using data from 15-30 minutes:

     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:

  • rmADAM10: 0.1 µg
  • Substrate: 10 µM

FAQs

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