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Recombinant Human ADAMTS13 (Full Length) Protein, CF

R&D Systems | Catalog # 6156-AD

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human ADAMTS13 (Full Length) Protein (6156-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

CHO

Accession Number

Q76LX8

Applications

Enzyme Activity
View all ADAMTS13 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human ADAMTS13 protein
Gln34-Thr1427, with a C-terminal 10-His tag

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Ala75, Gln34 inferred from enzymatic pyroglutamate treatment revealing Gln35

Predicted Molecular Mass

147 kDa & 151 kDa

SDS-PAGE

160-180 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, FRETS-VWF73.
The specific activity is >10 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6156-AD
Formulation Supplied as a 0.2 μm filtered solution in HEPES and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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 opening.

Background: ADAMTS13

ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin motifs 13), also known as von Willebrand Factor (vWF) cleaving protease, is a member of the family of secreted zinc proteases with a multidomain structure (1-3). The protein precursors consist of a signal peptide and following domains: pro, catalytic, disintegrin-like, thromobspondin type 1 (TSP1) motif, a cysteine-rich domain, a spacer region, a second set of seven TSP1 repeats, and two CUB domins. The only known substrate of ADAMTS13 is vWF, a blood glycoprotein with two homeostatic functions (4). It is required for platelet adhesion to sites of vascular damage and acts as a carrier protein for blood-clotting factor VIII in the circulation. It exists in plasma as multimers, the largest of which effectively mediate platelet adhesion. ADAMTS13 cleaves multimeric vWF in the A2 domain at the position between Tyr1605 and Met1606. A defect in ADAMTS13 activity is a cause of congenital thrombotic thrombocytopenic purpura (TTP), also known as Upshaw Schulman syndrome. Lack of ADAMTS13 activity allows unusually high concentrations vWF (UlvWF) to accumulate in plasma (5). These UlvWF multimers have a tendency to agglutinate circulating platelets at sites with high levels of shear stress to cause TTP. The recombinant human vWFA2 cleaving activity of recombinant human ADAMTS13 can be inhibited by 5 mM 1,10‑phenanthroline.

References

  1. Furlan, M. et al. (1996) Blood. 87:4223.
  2. Porter, S. et al. (2005) Biochem. J. 386:15.
  3. Chung, D. W. and J.E. Saddler (2004) in Handbook of Proteolytic Enzymes, Barret, A. J. et al. eds. pp. 747.
  4. Wu, J.J. et al. (2006) PNAS. 103:18470.
  5. Levy, G.G. et al. (2005) Blood. 106:11.

Long Name

A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 13

Alternate Names

TTP, vWF-CP

Entrez Gene IDs

11093 (Human); 279028 (Mouse); 362091 (Rat)

Gene Symbol

ADAMTS13

UniProt

Q76LX8

Additional ADAMTS13 Products

Product Documents for Recombinant Human ADAMTS13 (Full Length) Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human ADAMTS13 (Full Length) Protein, CF

For research use only

Citations for Recombinant Human ADAMTS13 (Full Length) Protein, CF

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Protocols

View specific protocols for Recombinant Human ADAMTS13 (Full Length) Protein, CF (6156-AD):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Human ADAMTS13 (rhADAMTS13) Full Length (Catalog # 6156-AD)
  • Substrate: FRETS-VWF73 (AnaSpec, Catalog # 63728), 100 µM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhADAMTS13 to 20 µg/mL in Assay Buffer.
  2. Dilute Substrate to 8 µM in Assay Buffer.
  3. Load 50 µL of dilute rhADAMTS13 into a plate, and start the reactions by adding 50 µL of 8 µM Substrate.  Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 8 µM Substrate.
  4. Read at excitation and emission wavelengths of 340 nm and 450 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 FRETS-25-STD1 (Peptides International, Catalog # STD-3720-V).

Per Well:
  • rhADAMTS13: 1 µg
  • Substrate: 4 µM

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