TAT Complex Paired Antibody Set
Affinity’s TAT Complex Paired Antibody Set consists of matched capture and detecting antibodies that have been titrated and optimized for use in sandwich style ELISA assays. The product as provided contains sufficient capture and detecting antibodies for five full 96-well microplates and contains a detailed protocol sheet containing directions for use, recipes for solutions and sources for additional materials required. This TAT Complex Paired Antibody Set is intended to facilitate the end user in establishing an “in-house” immunoassay for research purposes only and must not be used for diagnostic applications. Assay validation is the responsibility of the end user.
Product Code: TAT-EIA
- Capture Antibody (TAT-EIA-C): One yellow-capped vial containing 0.5 ml of polyclonal affinity purified anti-thrombin antibody for coating plates.
- Detecting Antibody (TAT-EIA-D): One red-capped tube containing 0.5 ml of peroxidase conjugated affinity-purified polyclonal anti-ATIII antibody for detection of captured TAT complex.
- Conjugate Diluent: Five neutral-capped tubes each containing 10 ml of universal diluent for peroxidase conjugated antibody. Each vial sufficient for one 96-well plate.
Species Cross Reactivity: View Chart
Product Datasheet: Thrombin Antithrombin Complex - TAT - Matched Pair Antibody Set for ELISA
Description of Thrombin Antithrombin Complex (TAT Complex)
The activation of coagulation ultimately leads to the activation of prothrombin to the enzyme thrombin. Unless regulated, thrombin will act on its natural substrates that include fibrinogen, factor V, factor VIII, factor XIII, Protein C, TAFI as well as specific receptors on platelets and endothelial cells. The activity of thrombin in plasma is regulated in part through interaction with protease inhibitors. Based on kinetic rates and physiological concentrations, the primary inhibitor of thrombin in plasma is antithrombin (ATIII), followed by heparin cofactor II and alpha-2-macroglobulin.
The thrombin-antithrombin complex (TAT) results when thrombin cleaves a scissile bond near the C-terminus of ATIII, forming a covalent, 1:1 acyl enzyme intermediate with ATIII with an apparent mass of 96 kDa. Calcium is not required for this interaction, but the rate of thrombin inhibition by ATIII can be accelerated 1000-fold by optimal concentrations of heparin. Although TAT complex is relatively stable, under conditions of elevated pH and/or enzyme excess, enzymatic degradation and even dissociation of the complex can occur. In serum, vitronectin (also known as S-Protein) has been reported to form larger ternary complexes with TAT (350 kDa), but these ternary S-TAT complexes are not covalently linked and are not stable to denaturants such as SDS. TAT complex is cleared from circulation by serpin-enzyme complex receptors on the surface of hepatocytes, with a half-life of 15 minutes1-4.
References and Reviews
- Bauer KA; Laboratory Markers of Coagulation Activation; Arch Pathol Lab Med 117, pp 71-77, 1993.
- Pelzer H, Schwarz A, Heimburger N; Determination of Thrombin-Antithrombin Complexes in Plasma with an Enzyme-Linked Immunosorbent Assay; Thrombosis and Haemostasis 59, pp 101-106, 1988.
- Griffith MJ, Lunblad RL; Dissociation of Antithrombin III-Thrombin Complex. Formation of Active and Inactive Antithrombin III; Biochemistry 20, pp 105, 1981.
- Preissner KT, Zwicker L, Muller-Berghaus G; Formation, characterization and detection of a ternary complex between S protein, thrombin and antithrombin III in serum; Biochem. J. 243, pp 105-111, 1987.