ADAMTS13, Recombinant, Human (A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif, 13)

ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin motifs 13), also known as von Willebran 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, cystein-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, Tyr 1605-Met 1606. 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 starts at the N-terminus of the pro domain and ends in the spacer domain.\n\nBased on N-terminal amino acid sequencing, the N-terminus (Q34) of the purified, secreted rhADAMTS13 was blocked. The 665 amino acid residue rhADAMTS13 predicts a molecular mass of 73kD and migrates as an approximately 90kD protein band in SDS-PAGE under reducing conditions.\n\nActivity:\nMeasured by its ability to cleave recombinant human von Willebrand factor A2 domain. Under the described conditions, 0.5ug of rhADAMTS13 can cleave > 50% of 5ug of rhvWF-A2.\nThe rhvWF-A2 cleaving activity can be inhibited by 5 mM 1,10-phenanthroline.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 20ug/ml\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, aliquot to avoid repeated freezing and thawing and freeze at -70 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for at least 12 months.

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SPECIFICATIONS

Catalog Number

A0859-61V

Size

20ug

Applications

WB

Form

Supplied as 0.2um filtered liquid in 25mM Acetate, 10mM CaCl2, 1M NaCl, pH 5.5

Purity

~90%, as determined by SDS-PAGE and visualized by silver stain. Endotoxin: < 1.0 EU per 1ug

References

1. Furlan, M. et al., 1996, Blood. 87:4223 - 4234. 2. Porter, S. et al., 2005, Biochem. J. 386:15 - 27. 3. Chung, D. W. and Saddler, J. E. 2004, in Handbook of Proteolytic Enzymes, Barret, A. J. et al. eds. pp 747 - 751. 4. Wu, J.-J. et al. 2006, PNAS. 103:18470 - 18474. 5. Levy, G. G. et al. 2005, Blood. 106:11 - 16.

Additional Info

Recognizes human ADAMTS13.