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Analytical balance

Company: Mettler Toledo
Catalog#: AB54-S
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Chromatographic Analysis for Targeted Metabolomics of Antioxidant and Flavor-Related Metabolites in Tomato
Author:
Date:
2021-03-05
[Abstract]  

Targeted metabolomics is a useful approach to evaluate crop breeding studies. Antioxidant and flavor-related traits are of increasing interest and are considered quality traits in tomato breeding. The present study presents chromatographic methods to study antioxidants (carotenoids, vitamin C, vitamin E, phenolic compounds, and glutathione) and flavor-related characters (sugars and organic acids) in tomato. Two different extraction methods (for polar and apolar entities) were applied to isolate the targeted compounds. The extraction methods developed in this work were time and cost-effective since no further purification was needed. Carotenoids, vitamin C, glutathione, and phenolic acids were analyzed by HPLC-PDA using a RP C18 column at an appropriate wavelength for each compound.

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[摘要]  [摘要]靶向代谢麦克风是评估作物育种研究的有用方法。抗氧化剂和与风味有关的性状受到越来越多的关注,被认为是番茄育种中的品质性状。本研究提供了色谱方法来研究番茄中的抗氧化剂(类胡萝卜素,维生素C,维生素E,酚类化合物和谷胱甘肽)和与风味相关的特征(糖和有机酸)。应用了两种不同的提取方法(针对极性和非极性实体)来分离目标化合物。由于不需要进一步纯化,因此这项工作中开发的提取方法既省时又经济。使用RP C18色谱柱,通过RP-C18色谱柱在适当波长下对每种化合物进行类胡萝卜素,维生素C,谷胱甘肽和酚酸的分析。用RP C18和NH2色谱柱通过HPLC分析维生素E和糖,并分别通过FLD和RI检测器进行检测。此外,MSTFA衍生化后,使用Rtx 5DA色谱柱使用GC-FID分析有机酸。结果,开发了定量重要植物代谢物的灵敏分析方法,并在本文中进行了描述。这些方法不仅适用于番茄,但都还表征其他物种的香味相关的和抗氧化的化合物。因此,这些协议可用于指导作物育种的选择。

[背景]代谢组学是其受到关注的其潜在的援助作物育种研究施加生化途径。在番茄育种中,由于消费者对提高作物品质的偏好,抗氧化剂和与风味有关的代谢产物都受到了越来越多的关注。人类无法自己合成抗氧化剂分子; ...

Relative Quantification of NaV1.1 Protein in Mouse Brains Using a Meso Scale Discovery-Electrochemiluminescence (MSD-ECL) Method
Author:
Date:
2021-02-05
[Abstract]  

Densitometric analysis is often used to quantify NaV1.1 protein on immunoblots, although the sensitivity and dilution linearity of the method are usually poor. Here we present a protocol for quantification of NaV1.1 in mouse brain tissues using a Meso Scale Discovery-Electrochemiluminescence (MSD-ECL) method. MSD-ECL is based on ELISA (enzyme-linked immunosorbent assay) and uses electrochemiluminescence to produce measurable signals. Two different antibodies are used in this assay to capture and detect NaV1.1 respectively in brain tissue lysate. The specificity of the antibodies is confirmed by Scn1a gene knock-out tissue. The calibration curve standards used in this assay were generated with mouse liver lysate spiked with mouse brain lysate, instead of using a recombinant protein. We

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[摘要]  [摘要]尽管该方法的灵敏度和稀释度线性通常较差,但经常使用光密度分析法定量免疫印迹上的Na V 1.1蛋白。这里瓦特E存在的Na进行定量的协议V在使用小鼠脑组织1.1细观量表发现-电化学发光(MSD-ECL)方法。MSD-ECL基于ELISA(酶联免疫吸附测定),并使用电化学发光产生可测量的信号。此测定法中使用了两种不同的抗体来捕获和检测Na V 1.1分别在脑组织中溶解。Scn1a基因敲除组织证实了抗体的特异性。此测定法中使用的校准曲线标准品是用掺有小鼠脑裂解液的小鼠肝裂解液而不是重组蛋白生成的。我们证明该方法是合格的,可用于特异性,准确度和精密度定量的小鼠脑组织中的Na V 1.1 。


[背景]的Na V 1.1,也称为电压门控钠通道的α亚基,I型,是一种跨膜通过对编码蛋白SCN1A基因(Meisler等人,2010 )。功能性Na V 1.1的表达降低会导致Dravet综合征(DS),这是一种严重的早发性癫痫性脑病(Dravet等,2005)。Na V 1.1在生物样品中的表达已用作DS的非临床药理生物标记物,可以使用免疫印迹的光密度分析进行测量。密度测定法通常不如标准免疫测定法准确和灵敏。此外,由于蛋白质序列的同源性,某些Na V 1.1抗体可能会与其他电压门控钠通道(VGSC)发生交叉反应,包括Na V 1.2,Na V 1.3和Na ...

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