Home / ADAM10 / Recombinant Mouse ADAM10 Protein, CF

Recombinant Mouse ADAM10 Protein, CF

Catalog #: 946-AD
3 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
946-AD-020
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse ADAM10 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 >5 pmol/min/µg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse ADAM10 protein
Thr215-Glu673, with a C-terminal 10-His tag
Accession #
O35598
N-terminal Sequence
Analysis
Thr215
Structure / Form
Recombinant Mouse ADAM10 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Predicted Molecular Mass
52 kDa
SDS-PAGE
60 kDa, reducing conditions

Product Datasheets

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946-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.

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.

Assay Procedure

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
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: 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
Entrez Gene IDs
102 (Human); 11487 (Mouse)
Alternate Names
a disintegrin and metalloprotease domain 10; a disintegrin and metalloproteinase domain 10; AD10; ADAM 10; ADAM metallopeptidase domain 10; ADAM10; CD156c antigen; CD156c; CDw156; disintegrin and metalloproteinase domain-containing protein 10; EC 3.4.24; HsT18717; kuz; Kuzbanian protein homolog; Kuzbanian; MADM; MADMEC 3.4.24.81; Mammalian disintegrin-metalloprotease

Citations for Recombinant Mouse ADAM10 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.

3 Citations: Showing 1 - 3
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  1. Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function
    Authors: Burmeister, M;Fraunenstein, A;Kahms, M;Arends, L;Gerwien, H;Deshpande, T;Kuhlmann, T;Gross, CC;Naik, VN;Wiendl, H;Klingauf, J;Meissner, F;Sorokin, L;
    Science advances
    Species: Mouse
    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. Targeting ADAM-mediated ligand cleavage to inhibit HER3 and EGFR pathways in non-small cell lung cancer.
    Authors: Zhou BB, Peyton M, He B, Liu C, Girard L, Caudler E, Lo Y, Baribaud F, Mikami I, Reguart N, Yang G, Li Y, Yao W, Vaddi K, Gazdar AF, Friedman SM, Jablons DM, Newton RC, Fridman JS, Minna JD, Scherle PA
    Cancer Cell, 2006-07-01;10(1):39-50.
    Species: Human
    Sample Types:
    Applications: Enzyme Assay

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