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Formic acid

甲酸

Company: Sigma-Aldrich
Catalog#: F0507
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Uptake Assays in Xenopus laevis Oocytes Using Liquid Chromatography-mass Spectrometry to Detect Transport Activity
Author:
Date:
2017-10-20
[Abstract]  Xenopus laevis oocytes are a widely used model system for characterization of heterologously expressed secondary active transporters. Historically, researchers have relied on detecting transport activity by measuring accumulation of radiolabeled substrates by scintillation counting or of fluorescently labelled substrates by spectrofluorometric quantification. These techniques are, however, limited to substrates that are available as radiolabeled isotopes or to when the substrate is fluorescent. This prompted us to develop a transport assay where we could in principle detect transport activity for any organic metabolite regardless of its availability as radiolabeled isotope or fluorescence properties.

In this protocol we describe the use of X. laevis oocytes ...
[摘要]  非洲爪蟾卵母细胞是用于表征异源表达的第二活性转运蛋白的广泛使用的模型系统。历史上,研究人员依靠通过闪烁计数或荧光标记的底物通过分光荧光定量测量放射性标记底物的积累来检测运输活动。然而,这些技术仅限于可用作放射性标记的同位素的底物或当底物是荧光时。这促使我们开发了一种运输测定法,我们可以原则上检测任何有机代谢物的转运活性,无论其作为放射性标记的同位素或荧光性质如何。

在这个协议中,我们描述了X的使用。卵母细胞作为表达次要活性转运蛋白的异源宿主,以及如何进行摄取测定,然后通过液相色谱 - 质谱(LC-MS)检测和定量运输的代谢物。我们已经成功地使用这种方法来鉴定和表征称为硫代葡萄糖苷和氰基葡糖苷的植物防御代谢物的转运蛋白(Jørgensen等人,2017),然而该方法可用于表征任何转运蛋白底物可以通过LC-MS检测。

【背景】来自非洲爪蟾蛙(Xenopus laevis)的卵母细胞是用于异源表达和表征膜蛋白(即,转运蛋白和通道)的完整表达系统。 X。卵母细胞表达少量内源性膜蛋白,具有低的背景转运活性。此外,来自植物的次要活性转运蛋白(Boorer等人,1992; Theodoulou和Miller,1995; Nour-Eldin等人,2006),动物(Sumikawa 1981); ...

Purification of N-coronafacoyl Phytotoxins from Streptomyces scabies
Author:
Date:
2017-04-05
[Abstract]  This procedure is used for large-scale purification of N-coronafacoyl phytotoxins that are produced by the potato common scab pathogen Streptomyces scabies. The procedure employs organic extraction of S. scabies culture supernatants under alternating basic and acidic conditions in order to preferentially isolate the phytotoxin - containing carboxylic acid fraction of the supernatant. Preparative thin layer chromatography and semi-preparative reverse phase - high performance liquid chromatography are then used to further purify the individual N-coronafacoyl phytotoxins of interest. [摘要]  该方法用于大规模纯化由马铃薯常见的痂病原体耻骨链球菌sc疮产生的N, - 海胆酰基植物毒素。 该方法采用有机萃取方法。 在交替的碱性和酸性条件下培养上清液以优先分离含有上清液的含有毒素的羧酸部分。 然后使用制备型薄层色谱法和半制备型反相 - 高效液相色谱法进一步纯化所需的单独的N, - 焦油酰甲酰植物毒素。

马铃薯普通痂病是由革兰氏阳性,丝状,链霉菌属的土壤细菌引起的经济上重要的作物病。第一个描述和最好的特征是痂病引起链霉菌属(Streptomyces spp。)是具有全球分布(Bignell等人,2010)的Streptomyces scabies (syn。 S。scabiei 。目前常见的痂病治理方法包括作物轮作,灌溉和土壤熏蒸;然而,这些策略经常失败,产生不一致的结果或者环境不友好(Dees和Wanner,2012)。为了开发更好的疾病控制策略,我们必须首先了解 S使用的分子机制。 sc疮感染植物并诱发疾病症状。研究表明,S的能力。引起疾病的sc疮是由于在感染过程中产生不同作用的毒力因子。在由s生产的已知或潜在的毒力因子中。 sc疮是植物毒素的家族,被称为具有类似于植物激素茉莉酸的可能起到抑制作用的 N - 角鲨烯酰植物毒素(也称为COR-样代谢产物)病原体感染期间的植物免疫应答(Bignell等人,2010; ...

Glucosinolates Determination in Tissues of Horseradish Plant
Author:
Date:
2015-08-20
[Abstract]  Glucosinolates (GLS) are secondary metabolites mainly found in plants belonging to the Brassicaceae family, including also horseradish (Armoracia rusticana G. Gaertn., B. Mey. & Scherb), a popular spice with a characteristic pungent flavor due to the abundance of GLS. Such compounds exhibit antibacterial, antifungal, and insecticidal activities, as well as human health properties. Therefore, it is very important to have a full understanding of their levels and profiles in plants. However, the characterization of GLS from horseradish crude extracts is a tough task, due to the complexity of the vegetal matrix and the occurrence of many GLS in trace amounts. Here we describe two alternative effective and rapid methods for GLS characterization in horseradish plants: Liquid chromatography ... [摘要]  硫代葡萄糖苷(GLS)是次要代谢物,主要在属于十字花科的植物中发现,包括辣根(Armoracia rusticana G.Gaertn,B.Mey。&Scherb),由于GLS丰度而具有特征刺激性风味 。 这种化合物表现出抗菌,抗真菌和杀虫活性以及人类健康性质。 因此,对其在植物中的水平和谱的完全理解是非常重要的。 然而,由于植物基质的复杂性和痕量的许多GLS的出现,来自辣根粗提取物的GLS的表征是艰难的任务。 在这里我们描述两种替代的有效和快速的方法在辣根植物的GLS表征:液相色谱耦合高分辨率质谱(LC-MS)测定完整的GLS和HPLC-UV测定脱硫-GLS。

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