Methods for removing contammatmg FAD and alkahne phosphatase from apo-ammoaad oxldase are described A method for phosphorylatmg flavm ademne dmucleotlde is also described Its hydrolysis to flavm ademne dmucleotlde catalysed by alkahne phosphatase IS demonstrated by hqmd chromatography Flavm ademne d
Fluorophotometric enzyme immunoassay of thyroid-stimulating hormone
โ Scribed by Nobumasa Kato; Hiroshi Naruse; Minoru Irie; Akio Tsuji
- Publisher
- Elsevier Science
- Year
- 1979
- Tongue
- English
- Weight
- 480 KB
- Volume
- 96
- Category
- Article
- ISSN
- 0003-2697
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โฆ Synopsis
A method for enzyme immunoassay of thyroid-stimulating hormone (TSH) is described. TSH was conjugated with horseradish peroxidase according to periodate oxidation method. Separation of the bound and free was obtained by double-antibody solid-phase technique using Sepharose 4B-anti-rabbit immunoglobulin G (IgG)-goat IgG. The fluorescence reaction using tyramine and hydrogen peroxide as substrates was used for the determination of enzyme activity in order to increase the sensitivity of enzyme immunoassay. The standard curve for serum TSH was satisfactory to recognize TSH concentrations as 0.06 ,uU/tube. TSH values obtained by this method correlated well with those obtained by radioimmunoassay (r, 0.96). The coefficients of variation were 1.8 to 5.3% (within assay) and 5.1 to 19.5% (between assay). The method is about equal to radioimmunoassay with respect to sensitivity. Since it requires minimal equipment and is less expensive than radioimmunoassay, it is possible to perform routine assays even in laboratories with limited facilities.
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