ルシゲニンを用いるインベルターゼ活性の化学発光測定法とその酵素イムノアッセイへの応用

DOI NDLデジタルコレクション Web Site Web Site 被引用文献3件

書誌事項

タイトル別名
  • Chemiluminescent assay for invertase activity by using lucigenin and its application to chemiluminescent enzyme immunoassay.
  • ルシゲニン オ モチイル インベルターゼ カッセイ ノ カガク ハッコウ ソク

この論文をさがす

抄録

A highly sensitive chemiluminescent assay for invertase activity by using lucigenin has been developed, and was applied to chemiluminescent enzyme immunoassay with invertase as label enzyme. Invertase is an enzyme that catalyses the hydrolysis of saccharose of form glucose and fructose. By adding lucigenin solution into the hydrolyzed solution, intensive light was produced. The procedure for the assay of invertase activity is as follows: the mixture of 0.1 ml of invertase 0.2 ml of 0.3M saccharose solution and 0.2 ml of 0.25 M acetate buffer (pH 5.0) was allowed to stand at 4°C, over night, and after 0.1 ml of 0.5 M phosphate buffer (pH 7.0) was added. The mixture was heated for 1 min in boiling water. Aliquot of 0.1 ml of this reaction mixture was assayed by adding lucigenin solution (1.2 × 10-3% lucigenin -1 × 10-3 M Triton X-100 -0.16 M KOH). The light intensity was measured with a luminometer (waiting time, 15s, integrating time, 1530 s). A log/log linear relationship between the chemiluminescence intensity and invertase activity was obtained from 102 to 104 μU/ assay, and the detection limit of invertase by this reaction was about 10-15 mol. The relative standard deviation of invertase was 1.75.9% (n=10). And a new chemiluminescent enzyme immunoassay of 17α-hydroxyprogesterone and thyroxine were developed by using invertase as label enzyme. The measurable ranges of 17α-hydroxyprogesterone and thyroxine were 10 to 500 pg and 25 to 500 pg/assay, respectively. The 17α-hydroxyprogesterone values in serum samples were assayed by both fluorescent and the present methods. The results showed good correlation ( Y (proposed enzyme immunoassay) = 0. 81X (fluorescent enzyme immunoassay) + 0. 11, r = O. 90, n = 30).

収録刊行物

  • 分析化学

    分析化学 37 (3), 123-127, 1988

    公益社団法人 日本分析化学会

被引用文献 (3)*注記

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ