Recombinant Human ADAMTS13 Protein, CF Summary
Product Specifications
Gln34 - Trp688
with a C-terminal 10-His tag
Analysis
Product Datasheets
4245-AD
Carrier Free
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.
4245-AD
| Formulation | Supplied as a 0.2 μm filtered solution in HEPES and NaCl. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
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) (Catalog # 4245-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
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).
- rhADAMTS13: 0.25 µg
- Substrate: 4 µM
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 multi-domain structure (1‑3). The protein precursors consist of a signal peptide and following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, spacer, 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, Tyr1605‑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 large vWF (UlvWF) to occur in plasma (5). These UlvWF multimers have tendency to agglutinate circulating platelets at sites with high levels of shear stress to cause TTP. The purified rhADAMTS13 ends in the spacer domain. The rhvWF‑A2 cleaving activity of rhADAMTS13 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. 747-751.
- Wu, J.J. et al. (2006) PNAS. 103:18470.
- Levy, G.G. et al. (2005) Blood. 106:11.
Citations for Recombinant Human ADAMTS13 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.
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Citations: Showing 1 - 5
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ADAMTS13 controls vascular remodeling by modifying VWF reactivity during stroke recovery
Authors: H Xu, Y Cao, X Yang, P Cai, L Kang, X Zhu, H Luo, L Lu, L Wei, X Bai, Y Zhu, BQ Zhao, W Fan
Blood, 2017-04-20;0(0):.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo -
Probing ADAMTS13 substrate specificity using phage display.
Authors: Desch, Karl C, Kretz, Colin, Yee, Andrew, Gildersleeve, Robert, Metzger, Kristin, Agrawal, Nidhi, Cheng, Jane, Ginsburg, David
PLoS ONE, 2015-04-07;10(4):e0122931.
Species: Human
Sample Types: Whole Cells
Applications: Enzyme Assay -
Plasmin cleavage of von Willebrand factor as an emergency bypass for ADAMTS13 deficiency in thrombotic microangiopathy.
Authors: Tersteeg C, de Maat S, De Meyer S, Smeets M, Barendrecht A, Roest M, Pasterkamp G, Fijnheer R, Vanhoorelbeke K, de Groot P, Maas C
Circulation, 2014-01-21;129(12):1320-31.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
The endogenous proteoglycan-degrading enzyme ADAMTS-4 promotes functional recovery after spinal cord injury.
Authors: Tauchi R, Imagama S, Natori T
J Neuroinflammation, 2012-03-15;9(0):53.
Species: Rat
Sample Types: Tissue Homogenates
Applications: Bioassay -
A novel calcium-binding site of von Willebrand factor A2 domain regulates its cleavage by ADAMTS13.
Authors: Zhou M, Dong X, Baldauf C, Chen H, Zhou Y, Springer TA, Luo X, Zhong C, Grater F, Ding J
Blood, 2011-03-08;117(17):4623-31.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay
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