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一种用于快速灵敏评估贻贝中冈田酸污染的蛋白磷酸酶2A抑制测定法。

A protein phosphatase 2A inhibition assay for a fast and sensitive assessment of okadaic acid contamination in mussels.

作者信息

Tubaro A, Florio C, Luxich E, Sosa S, Della Loggia R, Yasumoto T

机构信息

Dipartimento di Scienze Biomediche, Università di Trieste, Italy.

出版信息

Toxicon. 1996 Jul;34(7):743-52. doi: 10.1016/0041-0101(96)00027-x.

Abstract

The specific inhibitory activity exerted by okadaic acid on protein phosphatase 2A was used to assess the presence of okadaic acid in mussels, using a commercially available protein phosphatase 2A preparation. Under the conditions used, okadaic acid inhibits the enzymatic activity dose-dependently, with an IC50 = 0.26 ng/ml (0.32 nM). The assay is accurate and reproducible. Okadaic acid was detected in concentrations as low as 0.063 ng/ml in aqueous solutions and 2 ng/g in mussel digestive glands. Thirty naturally contaminated mussel samples were submitted to the protein phosphatase 2A inhibition assay as well as to an ELISA assay and to a MTT cytotoxicity assay, with similar results. The proposed assay is sensitive, rapid and does not require expensive equipment. These characteristics make it a good candidate for employment in the routine assessment of okadaic acid shellfish contamination.

摘要

利用冈田酸对蛋白磷酸酶2A的特异性抑制活性,使用市售的蛋白磷酸酶2A制剂来评估贻贝中冈田酸的存在情况。在所采用的条件下,冈田酸以剂量依赖方式抑制酶活性,IC50 = 0.26 ng/ml(0.32 nM)。该检测方法准确且可重复。在水溶液中检测到的冈田酸浓度低至0.063 ng/ml,在贻贝消化腺中的浓度为2 ng/g。30个自然污染的贻贝样本接受了蛋白磷酸酶2A抑制检测、酶联免疫吸附测定(ELISA)以及MTT细胞毒性检测,结果相似。所提出的检测方法灵敏、快速,且不需要昂贵的设备。这些特性使其成为常规评估贝类中冈田酸污染的良好选择。

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