Improved assay conditions for automated antithrombin IIi determinations with the chromogenic substrate S-2238

Van Voorthuizen, H.; Kluft, C.

Thrombosis and Haemostasis 52(3): 350-353

1984


ISSN/ISBN: 0340-6245
PMID: 6531759
Document Number: 236700
Assays for antithrombin III activity using the chromogenic substrate S-2238 were automated in various modifications on various instruments. Standard curves for the assay showed a deviating "0% point" in several but not all modifications. Addition of polyethylene glycol 6000/8000 (0.40-1.0%) minimizes this deviation. However, the deviation of the 0% point was not completely abolished in every modification of the test. Examination of Lineweaver-Burk plots showed nonlinearity at high concentrations of S-2238 as used in the antithrombin III assays. Addition of Tween-80 (0.01%) restores the Lineweaver-Burk plot and, at the same time, the standard curve in the assays is found to be linear. Therefore, we recommend the use of PEG 6000/8000 as well as Tween-80 in automated assay systems.

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