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优化用于巨噬细胞和血清的精氨酸酶活性微量测定法。

Optimization of the arginase activity assay micromethod for macrophages and sera.

机构信息

Laboratoire de Parasitologie, Institut Pasteur de Bangui, BP 923, Bangui, Central African Republic.

Laboratoire de Parasitologie, UMR 177 IRD/CIRAD "INTERTRYP," Université Bordeaux, Bordeaux, F-33000, France.

出版信息

BMC Res Notes. 2023 Aug 29;16(1):188. doi: 10.1186/s13104-023-06462-4.

Abstract

OBJECTIVE

We optimized the spectrophotometric micromethod for the determination of arginase activity based on the Corraliza et al. modification of Schimke's method. Arginase activity in sera from patients suffering from human African trypanosomiasis, in macrophage lysates from trypanosome-infected mice, and in purified bovine liver arginase was compared using the conventional and optimized micromethods.

RESULTS

The sensitivity of both micromethods was comparable. However, our optimized method has the following advantages: it uses small sample volumes (6 µl per assay vs. 50 µl) and reagent volumes (200 µl vs. 400 µl), it can be carried out in a single microplate well, thereby minimizing handling, and it requires fewer materials and utilizes readily available equipment. Our optimized method proved to be applicable and well suited for small-volume samples and resource-poor laboratories.

摘要

目的

我们对基于 Corraliza 等对 Schimke 方法的修改的分光光度微法定量测定精氨酸酶活性的方法进行了优化。使用常规和优化的微法定量测定了患有非洲人类锥虫病的患者的血清、感染锥虫的小鼠的巨噬细胞裂解物以及纯化的牛肝精氨酸酶中的精氨酸酶活性。

结果

两种微法定量测定的灵敏度相当。但是,我们的优化方法具有以下优点:它使用小样本量(每个测定 6 μl 与 50 μl)和试剂量(200 μl 与 400 μl),可以在单个微孔板孔中进行,从而最大程度地减少了操作,并减少了所需的材料和利用了现成的设备。我们的优化方法证明适用于小体积样本和资源匮乏的实验室。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c153/10466829/f114f225f7b2/13104_2023_6462_Fig1_HTML.jpg

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