A modified collagenase assay method based on the use of p-dioxane.
A modified collagenase assay method based on the use of p-dioxane.
复制标题
基于使用对二恶烷的改良胶原酶测定方法。
DOI：
10.1016/0003-2697(79)90635-3
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发表时间：
1979
影响因子：
2.9
通讯作者：
M. Nimni
中科院分区：
文献类型：
--
作者：
T. Ishikawa;M. Nimni
人皮肤成纤维细胞在单层培养中合成的放射性胶原被用作胶原酶的底物。该底物的高比活性（75，000 cpm/μg）和使用对二氧六环作为未消化胶原蛋白的沉淀剂允许在35°C和pH 7.5下用胶原蛋白溶液测定该酶。所用的稀释度足以防止胶原分子聚集，从而排除使用倾向于降低酶活性的凝胶形成抑制剂。使用1小时孵育，该方法在10 - 100 ng细菌胶原酶范围内具有重现性（SD ± 2.3%）和线性。脊椎动物胶原酶活性也很容易用这种方法测量。
Radioactive collagen synthesized by human skin fibroblasts in monolayer culture was used as a substrate for collagenase. The high specific activity of this substrate (75,000 cpm/μg) and the use of p-dioxane as a precipitant of the undigested collagen permit this enzyme to be assayed with collagen in solution at 35°C and pH 7.5. The dilutions used are sufficient to prevent the collagen molecules from aggregating, thus precluding the use of inhibitors of gel formation which tend to decrease the activity of the enzyme. Using a 1-h incubation, the procedure is reproducible (SD ± 2.3%) and linear over the range from 10 to 100 ng of bacterial collagenase. Vertebrate collagenase activity is also easily measured with this method.