A versatile assay method of antithrombin III using thrombin and Tos-Gly-Pro-Arg-Pna

Automation of Antithrombin III Methods on Routinely Available Instruments

“…The choice of an instrument should be directed by the number of tests performed per day, the size of the workload (small or large daily series, large weekly series), as well as tests performed on the same instrument (coagulation or biochemistry). Choice of instrument also depends on whether tests will be performed routinely and 36 Dettori et al 20 Andreasen 6 Holmes et al 28 Brandt et al 11 Bartl and Lill 8 Scully and Kakkar 39 Conard et al 19 Cederholm-Williams 13 Holmes et al 28 Aiach et al 4 Droullé et al 21 Kahle et al 30 Bartl and Lill 8 Dettori et al 20 Bartl and Lill 8 Prellwitz et al 38 Frantzen et al 24 Fareed et al 23 Prellwitz et al 38 whether the biologist wants to be able to modify conditions of the tests. The improvement of the quality of the results and the rapidity of the instruments should allow large-scale surveys of AT III, such as for epidemiologic purposes.…”

“…In contrast, chromogenic methods are easily automated by kinetic rate technique, and most of the instruments used in clinical chemistry laboratories are appropriate. Results obtained with the following instruments have been published: GEMSAEC centrifugal analyzer, 6,20,36 Multistat microcentrifugal analyzer, 28 ABA-100 Abbott, 8,11 Vitatron Automated Kinetic Enzyme and Substrate Analyzer, 8,30 Gilford 3500, 13,19,28 Isamat Isabiologie, 4 Kem-O-Mat 2T Coultronics, 21 LKB, 39 Olli 3000, 20 Eppendorf 5020, 8 Cobas Bio, 24,38 and aca DuPont. 17,23,38 The ABA-100 is only semiautomated.…”

“…Temperature of the reaction is frequently 37°C but may be 25°C, 8,30 30°C, 19,24,28 or room temperature. 4 Proportions of reagents vary.…”

Application of Several Chromogenic Substrate Assays to Automated Instrumentation for Coagulation Analysis

“…Methods for the Vitatron AKES, the Abbott ABA 1 00, and the Eppendorf 5020 Analyzer using this assay system have been described. 3 Applications to a variety of other systems have also been performed, for example, the CentrifiChem and Cobas Bio centrifugal analyzers, the Vitatron PA 800, the Eppendorf ACP 5040, and the LKB Mark II. Application to the Hitachi 705 system is described in detail as an example of the applicability of this assay to automated instruments.…”

Protease nexin-1 activity in cultured Schwann cells

“…This was done to prevent complex formation between PN-1 and proteases in the FCS. After removal from the cells the medium was concentrated 5-fold using Centricon filters (Amicon, Danvers, MA) and assayed for thrombin inhibitory activity in a Chromozym-TH (Boehringer Mann° heim, Indianapolis, IN) calorimetric assay [2]. Preincubation of the conditioned medium with anti-PN-1 antibody was used to test whether the thrombin inhibition was due to PN-1.…”