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PIPETMAN P20, 2 TO 20UL

P20移液器经典移液器

Company: Gilson
Catalog#: F123600
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Formalin Murine Model of Pain
Author:
Date:
2017-12-05
[Abstract]  Pain research is mostly based on experimental assays that use animal models, which may allow deciphering the physiopathology of this condition and to propel drug discovery. The formalin nociception test is considered one of the most predictive approaches to study acute pain in rodents. This test permits monitoring pain-related responses (i.e., itch) caused by a subcutaneous injection of an inflammatory agent, namely 2.5% formalin solution, in the hind paw. After the injection, two distinct time periods or phases of licking/biting behaviour occur, which are separated by a quiescent period. Importantly, these phases differ in duration and underlying mechanisms. Hence, the initial acute phase (phase I), commonly recorded for 5 min just after formalin administration, reflects acute ... [摘要]  疼痛研究主要基于使用动物模型的实验分析,这可能允许破译这种病症的病理生理学并推动药物发现。福尔马林伤害感受测试被认为是研究啮齿动物急性疼痛的最有预见性的方法之一。该测试允许监测由后部皮下注射炎症剂,即2.5%福尔马林溶液引起的疼痛相关反应(即,瘙痒)。注射后,发生两个不同的舔/咬行为的时间段或阶段,这些时段或阶段由静止期隔开。重要的是,这些阶段的持续时间和基本机制不同。因此,通常在福尔马林给药后5分钟记录的初始急性期(阶段I)反映了急性外周疼痛,可能是由于通过TRPA1通道直接激活伤害感受器。另一方面,在静止期(5-15分钟)后开始并且通常记录15-30分钟的阶段II是由于正在进行的炎症输入和中枢伤害性敏化。在这里,我们详细描述了用于在小鼠中进行可靠和可重复的福尔马林测试的方案。

【背景】福尔马林试验是允许确定小鼠伤害行为的实验分析。因此,通常在足底后爪(Tjolsen等人,1992)中皮下注射福尔马林后评估小鼠反应(即,舔and和退缩)。该试验最初是在七十年代后期描述的(Dubuisson和Dennis,1977),最初是由一只老鼠或一只猫的前爪背面注射50μl5%福尔马林组成的。此后,福尔马林试验已经被广泛用于评估伤害感受和炎症相关反应,因此根据每项研究的目的进行调整。小鼠和大鼠由于天生的梳理行为而经常使用(即,前爪舔)。因此,注射到啮齿动物后爪中足底表面的2.5%福尔马林(在生理盐水溶液中)通常用于诱导伤害性反应(即,爪,'退缩'和舔)持续45-90分钟(Tjolsen等人,1992)。 ...

Trimolecular Fluorescence Complementation (TriFC) Assay for Direct Visualization of RNA-Protein Interaction in planta
Author:
Date:
2017-10-20
[Abstract]  RNA-Protein interactions play important roles in various eukaryotic biological processes. Molecular imaging of subcellular localization of RNA/protein complexes in plants is critical for understanding these interactions. However, methods to image RNA-Protein interactions in living plants have not yet been developed until now. Recently, we have developed a trimolecular fluorescence complementation (TriFC) system for in vivo visualization of RNA-Protein interaction by transient expression in tobacco leaves. In this method, we combined conventional bimolecular fluorescence complementation (BiFC) system with MS2 system (phage MS2 coat protein [MCP] and its binding RNA sequence [MS2 sequence]) (Schonberger et al., 2012). Target RNA is tagged with 6xMS2 and MCP and RNA binding ... [摘要]  RNA-蛋白质相互作用在各种真核生物过程中起重要作用。 RNA /蛋白质复合物在植物中亚细胞定位的分子成像对于理解这些相互作用至关重要。然而,到目前为止,尚未开发在活植物中形成RNA-蛋白质相互作用的方法。最近,我们开发了一种三分子荧光互补(TriFC)系统,用于在烟草叶中瞬时表达的RNA-蛋白质相互作用的体内可视化。在这种方法中,我们将传统的双分子荧光互补(BiFC)系统与MS2系统(噬菌体MS2外壳蛋白[MCP]及其结合RNA序列[MS2序列])(Schonberger等人,2012)相结合, 。目标RNA用6xMS2标记,MCP和RNA结合蛋白与YFP片段融合。编码这种融合RNA和蛋白质的DNA构建体用土壤杆菌悬浮液渗入烟草叶中。通过共焦显微镜观察体内的RNA-蛋白质相互作用
【背景】近来,多种类型的长非编码RNA(lncRNA)已经被鉴定并显示出在转录调节和染色质修饰中起重要作用(St Laurent等人,2015)。到目前为止,lncRNA介导的功能的大多数分子机制与RNA-蛋白质相互作用密切相关(St ...

NP-40 Fractionation and Nucleic Acid Extraction in Mammalian Cells
Author:
Date:
2017-10-20
[Abstract]  This technique allows for efficient, highly purified cytoplasmic and nuclear-associated compartment fractionation utilizing NP-40 detergent in mammalian cells. The nuclear membrane is not disturbed during the fractionation thus leaving all nuclear and perinuclear associated components in the nuclear fraction. This protocol has been modified from Sambrook and Russell (2001) in order to downscale the amount of cells needed. To determine the efficiency of fractionation, we recommend using qPCR to compare the subcellular compartments that have been purified with equivalent amount of control whole cell extracts. [摘要]  该技术允许在哺乳动物细胞中利用NP-40洗涤剂进行高效,高度纯化的细胞质和核相关的分室分离。 在分离过程中核膜不受干扰,从而使核部分中的所有核和核周相关成分留下。 该协议已经从Sambrook和Russell(2001)修改,以便缩减所需的细胞数量。 为了确定分馏的效率,我们建议使用qPCR来比较已经用等量的对照全细胞提取物纯化的亚细胞室。
【背景】为了充分获得对细胞过程的理解,需要分离核和细胞质隔室。 有许多协议,甚至一些商业套件可用于帮助分离两个隔间。 然而,大多数需要高离心速度,产量差异甚至验证最终产品中的污染物量的方法也是很高的。 我们的协议在低速下使用小型台式离心机,以获得高纯度的细胞质提取物和核/核周组合相关隔室,以及数据分析,以验证污染物的百分比。 迄今为止,已经测试的细胞系是293T,HeLa和GHOST细胞系。 (Galvis,2014; Galvis等,,2014)。

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