Recombinant Human ADAMTS13 (Full Length) Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human ADAMTS13 (Full Length) Protein, CF Summary

Details of Functionality
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.
Source
Chinese Hamster Ovary cell line, CHO-derived human ADAMTS13 protein
Gln34-Thr1427, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Ala75, Gln34 inferred from enzymatic pyroglutamate treatment revealing Gln35
Protein/Peptide Type
Recombinant Enzymes
Gene
ADAMTS13
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
147 kDa & 151 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
160-180 kDa, reducing conditions
Publications
Read Publications using
6156-AD in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES and NaCl.
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • 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 4 µM Substrate.  Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 4 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human ADAMTS13 (Full Length) Protein, CF

  • A disintegrin and metalloproteinase with thrombospondin motifs 13
  • a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 13
  • ADAM metallopeptidase with thrombospondin type 1 motif, 13
  • ADAMTS13
  • ADAM-TS13
  • ADAMTS-13
  • DKFZp434C2322
  • EC 3.4.24.14
  • EC 3.4.24.82
  • EC 3.4.24.87
  • FLJ42993
  • MGC118899
  • MGC118900
  • TTP
  • TTPADAM-TS 13
  • vWF-cleaving protease
  • vWF-CP
  • vWF-CPC9orf8
  • VWFCPvon Willebrand factor-cleaving protease

Background

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.
  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.

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Bioinformatics

Gene Symbol ADAMTS13
Uniprot