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Mineral oil

矿物油

Company: Sigma-Aldrich
Catalog#: M5904
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Xenopus laevis Oocytes Preparation for in-Cell EPR Spectroscopy
Author:
Date:
2018-04-05
[Abstract]  One of the most exciting perspectives for studying bio-macromolecules comes from the emerging field of in-cell spectroscopy, which enables to determine the structure and dynamics of bio-macromolecules in the cell. In-cell electron paramagnetic resonance (EPR) spectroscopy in combination with micro-injection of bio-macromolecules into Xenopus laevis oocytes is ideally suited for this purpose. Xenopus laevis oocytes are a commonly used eukaryotic cell model in different fields of biology, such as cell- and development-biology. For in-cell EPR, the bio-macromolecules of interest are microinjected into the Xenopus laevis oocytes upon site-directed spin labeling. The sample solution is filled into a thin glass capillary by means of Nanoliter Injector and after that ... [摘要]  研究生物大分子的最令人兴奋的观点之一来自于新兴的细胞内光谱学领域,它能够确定细胞中生物大分子的结构和动力学。细胞内电子顺磁共振(EPR)光谱结合将生物大分子微注射到非洲爪蟾卵母细胞中非常适合于此目的。非洲爪蟾卵母细胞是生物学不同领域常用的真核细胞模型,如细胞和发育生物学。对于细胞内EPR,感兴趣的生物大分子通过定点自旋标记显微注射到非洲爪蟾卵母细胞中。通过Nanoliter注射器将样品溶液填充到薄玻璃毛细管中,然后通过小心地穿刺薄膜将其微注射入非洲爪蟾卵母细胞的黑色动物部分。之后,取决于最终的细胞内EPR实验的种类,将三个或五个显微注射的非洲爪蟾卵母细胞装载到Q波段EPR样品管中,随后进行任选的休克冷冻(用于实验冷冻溶液)并且在期望的温育时间之后测量(在低温或生理温度下)。由于显微注射样品的细胞毒性作用和顺磁性自旋标记在还原性细胞环境中的稳定性,孵育时间受到限制。通过监测细胞形态和减少动力学来量化这两个方面。

【背景】电子顺磁共振(EPR)光谱学是用于表征顺磁系统的选择方法(Atherton,1993; Gerson等人,1994; Jeschke和Schweiger,2001)。反磁性生物大分子可以通过定点自旋标记(SDSL)进行EPR光谱学分析,通常使用氮氧化物作为自旋标记(Hubbell和Altenbach,1994; Feix和Klug,2002; ...

Two-electrode Voltage-clamp Recordings in Xenopus laevis Oocytes:Reconstitution of Abscisic Acid Activation of SLAC1 Anion Channel via PYL9 ABA Receptor
Author:
Date:
2017-01-20
[Abstract]  Two-Electrode Voltage-Clamp (TEVC) recording in Xenopus laevis oocytes provides a powerful method to investigate the functions and regulation of ion channel proteins. This approach provides a well-known tool to characterize ion channels or transporters expressed in Xenopus laevis oocytes. The plasma membrane of the oocyte is impaled by two microelectrodes, one for voltage sensing and the other one for current injection. Here we list a protocol that allows robust reconstitution of multi-component signaling pathways. This protocol has been used to study plant ion channels, including the SLAC1 channel (SLOW ANION CHANNEL-ASSOCIATED 1), in particular SLAC1 activation by either the protein kinase OST1 (OPEN STOMATA 1), Ca2+-dependent protein kinases (CPKs) or the ... [摘要]  在非洲爪蟾卵母细胞中记录的双电极电压钳(TEVC)提供了一种强大的方法来研究离子通道蛋白的功能和调节。这种方法提供了一种众所周知的用于表征在非洲爪蟾卵母细胞中表达的离子通道或转运蛋白的工具。卵母细胞的质膜由两个微电极引起,一个用于电压感测,另一个用于电流注入。在这里,我们列出了一个允许多组分信号通路强制重组的协议。该方案已经用于研究植物离子通道,包括SLAC1通道(SLOW ANION CHANNEL-ASSOCIATED 1),特别是SLAC1通过蛋白激酶OST1(OPEN STOMATA 1),Ca 2 +依赖性蛋白激酶(CPK)或GHR1(GUARD细胞过氧化氢抗性1)跨膜受体样蛋白。显示了通过“单体”ABA(脱落酸)受体RCAR1 / PYL9(PYRABACT INRESISTANCE1 [PYR1] / PYR1-样[PYL] / ABA受体[RCAR]的调节因子]重建SLAC1阴离子通道的脱落酸活化的数据。通过共表达脱落酸信号核心的四个组分。该方案也适用于研究其他离子通道功能和调节机制,以及转运蛋白。背景 ...

Semi-thin Sectioning, Light and Fluorescence Microscopy of Floral Bud to Study Microspore Development in Arabidopsis
Author:
Date:
2016-03-05
[Abstract]  Pollen grains are male gametophytes produced within the pollen sacs of the anthers of the flower. Recent genetic studies have revealed several components involved in microspore development (Borg et al., 2009; Berger and Twell, 2011), and yet many components controlling microspore development remain to be identified. Semi-thin sectioning of anthers and light and fluorescence microscopy of floral bud (Kim et al., 2015) are the initial key experiments to characterize Arabidopsis mutants and transgenic plants for understanding the roles of new genetic components during microspore development. Herein, we describe a protocol for semi-thin sectioning of anthers and light and fluorescence microscopy of floral bud in Arabidopsis. [摘要]  花粉粒是在花的花药的花粉囊内产生的雄性配子体。 最近的遗传研究揭示了涉及小孢子发育的几种组分(Borg等人,2009; Berger和Twell,2011),然而仍有许多控制小孢子发育的组分被鉴定。 花药的半薄切片和花芽的光和荧光显微镜检查(Kim等人,2015)是表征拟南芥突变体和转基因植物的初始关键实验 在小孢子发育过程中新遗传组分的作用。 在这里,我们描述了拟南芥花药半花瓣和光和荧光显微镜的半薄切片的协议。

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