A sensitive colorimetric assay for thrombin, prothrombin and antithrombin III in human plasma using a new synthetic substrate
| Autor: | Hitomi Yuji, Niinobe Michio, Kanda Takao, Fujii Setsuro |
|---|---|
| Rok vydání: | 1982 |
| Předmět: |
Antithrombin III
Clinical Biochemistry Biochemistry Cofactor chemistry.chemical_compound Thrombin medicine Humans Zymography Fibrinolysin biology Activator (genetics) Hydrolysis Factor X Biochemistry (medical) Antithrombin Substrate (chemistry) General Medicine Heparin Electrophoresis Disc Kinetics chemistry biology.protein Colorimetry Prothrombin Oligopeptides Peptide Hydrolases circulatory and respiratory physiology medicine.drug |
| Zdroj: | Clinica Chimica Acta. 119:157-164 |
| ISSN: | 0009-8981 |
| DOI: | 10.1016/0009-8981(82)90328-x |
| Popis: | Benzoyl-L-leucyl-L-alanyl-L-arginine-alpha-naphthylester (Bz-Leu-Ala-Arg-NE) was synthesized as a new substrate for use in the assay of thrombin. In the assay alpha-naphthol released by the enzyme reaction was measured colorimetrically. With Bz-Leu-Ala-Arg-NE as substrate, the minimum detectable concentration of human thrombin was 0.0025 U. This assay using Bz-Leu-Ala-Arg-NE is a highly sensitive method for detecting prothrombin, thrombin and antithrombin III in human plasma. Prothrombin could be determined with 0.2 microliter of human plasma using Echis carinatus venom (ECV) as activator. Antithrombin III activity could be determined with 2 microliter of human plasma using human thrombin and heparin as cofactor. A zymogram of human prothrombin was prepared with Bz-Leu-Ala-Arg-NE as substrate. The preparation gave one band (pI 4.9) on polyacrylamide disc gel isoelectrophoresis. |
| Databáze: | OpenAIRE |
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