DiaPharma Group, Inc.

For In Vitro Research Use Only.

Chromogenic ELISA for the determination of ADAMTS-13 activity in human plasma. ADAMTS-13 is an enzyme that specifically cleaves unusually large von Willebrand Factor multimers, which induce platelet thrombus formation under high shear stress. If the activity of ADAMTS-13 is lowered for some reason, unusually large vWF multimers may accumulate within blood causing thrombosis due to platelet aggregation, which in turn may lead to TTP (Thrombotic thrombocytopenic purpura).

Advantages

1. ADAMTS-13 activity sensitivity down to 0.5% of normals
2. Sensitivity of measurement for inhibitors
3. No need for any special technique or instrument, just standard ELISA equipment
4. Short assay time (3h)
5. Calibrators and controls included

Assay Principle

Reagents -- Composition, Storage and Stability

1. ELISA test strips (12), with 8 wells each, coated with a monoclonal anti GST- Antibody. The drying agent is supplied in an aluminium bag. Stability after reconstitution: expiry date.
2. GST-vWF73 Substrate; 2 vials; lyophilized; 6 mL. Stability after reconstitution: 6 weeks.
3. Calibrators (Standards) numbered from 1 to 5; lyophilized; 1 vial each; 0.5mL. Stability after reconstitution: 6 months. Concentrations are lot-specific; consult the label on the vial.
4. High and low control plasma, lyophilized, 1 vial each, 0.5 mL. Stability after reconstitution: 6 months. Concentrations are lot-specific; consult the label on the vial.
5. Reaction buffer; 1 vial; 30 mL; ready to use. Stability after reconstitution: 6 months.
6. Conjugate: HRP conjugated monoclonal anti-N10 Antibody: 1 vial; 12mL; ready to use. Stability after reconstitution: 1 month.
7. Color reagent TMB (Tetramethylbenzidine); 1 vial; 12mL; ready to use. Stability after reconstitution: expiry date.
8. Wash Buffer concentrate, 10-fold concentrated, 1 vial, 32mL. Stability after reconstitution: 6 months.
9. Stop solution; sulphuric acid 0.5mol/L, 1 vial, 12 mL; ready to use. Stability after reconstitution: expiry date.
10. Sample dilution Microplate, 1 plate (ONLY for sample dilution!).

The expiry date printed on the labels applies to storage of the unopened vial at 2-8 °C.

Abstracts

• PMID: 17562615. Plasma levels of ADAMTS13 antigen determined with an enzyme immunoassay using a neutralizing monoclonal antibody parallel ADAMTS13 activity Levels. Int J Hematol. 2007 Jun;85(5):403-7.
• PMID: 16934083. Novel monoclonal antibody-based enzyme immunoassay for determining plasma levels of ADAMTS13 activity. Transfusion. 2006 Aug;46(8):1444-52.