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＜departmental bulletin paper＞
A Simple, Easy and Turbidimetric Assay of Thrombolytic Enzyme Activity in Microtiter Plates

Creator
Author PID
K001564
Creator Name
原, 敏夫
Hara, Toshio
Affiliation
Affiliation Name
九州大学農学部遺伝子資源研究センター微生物遺伝子部門
Microbial Genetics Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University
Creator Name
田所, 優子
Tadokoro, Yuko
Affiliation
Affiliation Name
九州大学農学部遺伝子資源研究センター微生物遺伝子部門
Microbial Genetics Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University
Creator Name
樽井, 寛
Tarui, Hiroshi
Affiliation
Affiliation Name
九州大学農学部遺伝子資源研究センター微生物遺伝子部門
Microbial Genetics Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University
Creator Name
里山, 俊哉
Satoyama, Toshiya
Affiliation
Affiliation Name
九州大学農学部遺伝子資源研究センター微生物遺伝子部門
Microbial Genetics Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University
Language
Japanese
Publisher
九州大學農學部
Faculty of Agriculture, Kyushu University
Date
1995-11
Source Title
Science bulletin of the Faculty of Agriculture, Kyushu University
Vol
50
Issue
1/2
First Page
31
Last Page
35
Publication Type
Version of Record
Access Rights
open access
JaLC DOI
https://doi.org/10.15017/23556
Related DOI
九州大學農學部學藝雜誌 || 50(1/2) || p31-35
Science bulletin of the Faculty of Agriculture, Kyushu University || 50(1/2) || p31-35
http://www.agr.kyushu-u.ac.jp/
Related URI
九州大學農學部學藝雜誌 || 50(1/2) || p31-35
Science bulletin of the Faculty of Agriculture, Kyushu University || 50(1/2) || p31-35
http://www.agr.kyushu-u.ac.jp/
Relation
九州大學農學部學藝雜誌 || 50(1/2) || p31-35
Science bulletin of the Faculty of Agriculture, Kyushu University || 50(1/2) || p31-35
http://www.agr.kyushu-u.ac.jp/

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Details

Record ID
23556
Peer-Reviewed
Unrefereed
Subject Terms
マイクロプレート
血栓溶解
酵素活性
濁度測定
フィブリン平板法
microtiter plates
thrombolysis
enzyme activity
turbidimetry
fibrin plate test
ISSN
0368-6264
NCID
AN0005519X
Notes
A turbidimetric assay for thrombolytic enzyme was performed in microtiter assay plates, wherein decrease in light scatter at 655 nm of fibrin plates in individual wells at 37℃ was recorded automatically at different times and used as a quantitative measure of fibrin dissolution. Use of an automated microtiter plate reader facilitated this assay considerably and allowed many assays to be performed rapidly, with a large amount of replication. The reliavility of the –ΔOD_<655>, CV%, caluculated by subtracting OD_<655> at individual indicated incubation time in eight microtiter wells from OD_<655> of microtiter wells before adding “natto” extract was found to be in 10% throughtout various degrees of dilution of “natto” extract. Logarithmic values of the degree of dilution of “natto” extract and –ΔOD_<655>/hr showed a linear relation, irrespective of the sample quality, indicating that this is a good method for the determination of thrombolytic activity in “natto” products.
1 )マイクロプレートとプレートリーダーを組み合わせた血栓溶解活性測定法を開発した. 本法は4時間以内に酵素活性を測定でき, 操作も簡便で, さらに基質量もきわめて少量ですみ, 低コストであった. 2 )生理的食塩水で市販納豆から抽出した酵素液で人工血栓は溶解され, 各反応時間後の吸光度を反応開始時の吸光度から差し引いて得られた-ΔOD_<655>値のCV%は, 用いたマイクロプレートの8連のウエルで10%以内となり, 高い信頼度を示した. 3 )-ΔOD_<655>を経時的にプロットすると測定時間内で直線性を示した. また, 単位時間当たりの-ΔOD_<655>の変化率から得られる直線の勾配と粗酵素であっても納豆から調製した血栓溶解酵素濃度の対数値がよく相関した. 4 )各種市販納豆中の血栓溶解酵素活性を測定したところ納豆により大きな違いのあることが分かった. このことより, 本法は血栓溶解酵素活性の測定に適した簡便な方法であることが明らかになった.
Created Date 2012.07.28
Modified Date 2022.11.11

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