Recombinant Human ADAMTS13 (Full Length) Protein, CF Summary
Gln34-Thr1427, with a C-terminal 10-His tag
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.
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.
|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.
- 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
- Dilute rhADAMTS13 to 20 µg/mL in Assay Buffer.
- Dilute Substrate to 8 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 340 nm and 450 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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).
- rhADAMTS13: 1 µg
- Substrate: 4 µM
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.
- Furlan, M. et al. (1996) Blood. 87:4223.
- Porter, S. et al. (2005) Biochem. J. 386:15.
- Chung, D. W. and J.E. Saddler (2004) in Handbook of Proteolytic Enzymes, Barret, A. J. et al. eds. pp. 747.
- Wu, J.J. et al. (2006) PNAS. 103:18470.
- Levy, G.G. et al. (2005) Blood. 106:11.
Citations for Recombinant Human ADAMTS13 (Full Length) 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.
Citations: Showing 1 - 6
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CaMKII orchestrates endoplasmic reticulum stress and apoptosis in doxorubicin-induced cardiotoxicity by regulating the IRE1alpha/XBP1s pathway
Authors: L Kong, Y Zhang, J Ning, C Xu, Z Wang, J Yang, L Yang
Journal of Cellular and Molecular Medicine, 2022-09-16;0(0):.
Distinct impact of IgG subclass on autoantibody pathogenicity in different IgG4-mediated diseases
Authors: Y Bi, J Su, S Zhou, Y Zhao, Y Zhang, H Zhang, M Liu, A Zhou, J Xu, M Pan, Y Zhao, F Li
Sample Types: Cell Culture Supernates
Applications: ELISA Standard
Optimization of anti-ADAMTS13 antibodies for the treatment of ADAMTS13-related bleeding disorder in patients receiving circulatory assist device support
Authors: T Ito, T Minamitani, M Hayakawa, R Otsubo, H Akiba, K Tsumoto, M Matsumoto, T Yasui
Scientific Reports, 2021-11-16;11(1):22341.
Sample Types: Protein
Applications: ELISA Capture, Surface Plasmon Resonance
High throughput protease profiling comprehensively defines active site specificity for thrombin and ADAMTS13
Authors: CA Kretz, K Tomberg, A Van Esbroe, A Yee, D Ginsburg
Sci Rep, 2018-02-12;8(1):2788.
ADAMTS13 and its variants promote angiogenesis via upregulation of VEGF and VEGFR2.
Authors: Lee, Manfai, Keener, Justin, Xiao, Juan, Long Zheng, X, Rodgers, George M
Cell Mol Life Sci, 2014-06-21;72(2):349-56.
Sample Types: Whole Cells
Effect of ADAMTS-13 on cerebrovascular microthrombosis and neuronal injury after experimental subarachnoid hemorrhage.
Authors: Muroi C, Fujioka M, Mishima K, Irie K, Fujimura Y, Nakano T, Fandino J, Keller E, Iwasaki K, Fujiwara M
J Thromb Haemost, 2014-04-01;12(4):505-14.
Sample Types: In Vivo
Applications: In Vivo
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