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Dimethyl sulfoxide

二甲亚砜(DMSO)

Company: Sigma-Aldrich
Catalog#: D8418
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Increasing the Membrane Permeability of a Fern with DMSO
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Date:
2018-06-20
[Abstract]  Cell membrane prevents the entrance of extra molecules (e.g., transcription and translation inhibitors) into the cell. For studying the physiological effects of transcription and translation inhibitors on Hymenophyllum caudiculatum fronds, we incubate fronds with 0.1% DMSO to test if this increases cell membrane permeability relative to incubation with ultrapure water. The study showed that DMSO could significantly improve the cell membrane permeability of filmy fronds. [摘要]  细胞膜防止额外分子(例如,转录和翻译抑制剂)进入细胞。 为了研究转录和翻译抑制剂对Hymenophyllum caudiculatum叶的生理影响,我们将叶用0.1%DMSO温育以测试它是否相对于用超纯水孵育增加细胞膜渗透性。 该研究表明DMSO可显着提高膜状叶的细胞膜透性。

【背景】薄膜蕨类植物最显着的特征之一是,除了维管组织外,叶子由单细胞层组成,缺少气孔。 在我们以前的工作中(Garcéset al。 2018年),我们用放线菌酮或放线菌素D孵育了Hymenophyllum caudiculatum叶,分别研究翻译或转录抑制的作用。 如果与薄膜蕨类植物一起孵育的外源抑制剂不影响植物生理,可能是因为抑制剂不能进入细胞。 为了改善抑制剂进入细胞,我们测试了0.1%DMSO水溶液是否对细胞膜通透性有影响,观察到碘化丙啶(PI)进入细胞。

In vitro Analysis of Ubiquitin-like Protein Modification in Archaea
Author:
Date:
2018-05-20
[Abstract]  The ubiquitin-like (Ubl) protein is widely distributed in Archaea and involved in many cellular pathways. A well-established method to reconstitute archaeal Ubl protein conjugation in vitro is important to better understand the process of archaeal Ubl protein modification. This protocol describes the in vitro reconstitution of Ubl protein modification and following analysis of this modification in Haloferax volcanii, a halophilic archaeon serving as the model organism. [摘要]  泛素样(Ubl)蛋白广泛分布于古细菌中并参与许多细胞途径。 为了更好地理解古细菌Ub1蛋白质修饰的过程,重建体外古细菌Ubl蛋白质缀合物的完善方法是很重要的。 该协议描述了Ubl蛋白质修饰的体外重建以及在作为模型生物的嗜盐古细菌Haloferax volcanii 中对这种修饰进行分析。

【背景】泛素(Ub)与靶蛋白共价连接的过程被称为泛素化,其控制真核细胞中大量的细胞过程(Glickman和Ciechanover,2002; Komander和Rape,2012)。遍在蛋白化由一系列酶(包括Ub激活酶(E1),Ub结合酶(E2s)和Ub连接酶(E3s))催化。泛素化的体外重建是确定酶之间或E3与蛋白质底物之间特异性的有用测定法(Zhao等人,2012)。在古细菌中,Ubl蛋白SAMP采用Ub折叠,并且与E1样酶UbaA催化的蛋白靶标异肽连接[Maupin-Furlow,(2014)综述]。尽管E1同系物在古细菌中广泛存在,但基于一级序列比较,在大多数古细菌中未预测经典E2或E3酶。我们最近对Haloferax volcanii的研究表明甲硫氨酸亚砜还原酶A(MsrA)是Ubl蛋白质修饰(sampylation)与UbaA一起在体内温和的氧化条件下和< (体外)(fu="">

Cobblestone Area-forming Cell Assay of Mouse Bone Marrow Hematopoietic Stem Cells
Author:
Date:
2018-05-05
[Abstract]  Bone Marrow Hematopoietic Stem Cells (HSCs) require bone marrow microenvironment for their maintenance and proliferation. Culture of Bone Marrow Mesenchymal Stromal Cells (MSCs) provides appropriate environmental signals for HSCs survival in vitro. Here, we provide a detailed protocol that describes culture conditions for MSCs, flow cytometric isolation of HSCs from mouse bone marrow, and perform co-culture of MSCs and HSCs known as Cobblestone area-forming cell (CAFC) assay. Altogether, CAFC assays can be used as a high-throughput in vitro screening model where efforts are made to understand and develop therapies for complex bone marrow diseases. This protocol needs 3 to 4 weeks starting from culturing MSCs, isolating LSK cells (HSCs), and to performing limited dilution ... [摘要]  骨髓造血干细胞(HSC)需要骨髓微环境来维持和增殖。 骨髓间充质基质细胞(MSC)的培养为体外HSC存活提供适当的环境信号。 在这里,我们提供了描述MSCs培养条件的详细方案,从小鼠骨髓中流式细胞术分离HSCs,并进行称为鹅卵石区域形成细胞(CAFC)分析的MSC和HSC的共培养。 总而言之,CAFC分析可用作高通量体外筛选模型,其中努力了解和开发复杂骨髓疾病的治疗方法。 该方案需要培养MSC,分离LSK细胞(HSC)和执行有限稀释CAFC测定3至4周。

【背景】HSC的增殖,存活和分化潜力非常依赖于其微环境,也被称为小生境。骨髓MSC支持HSC以使其在骨髓龛中保持静止状态。由生态位接收的内在和外在信号有助于将HSC分化为也称为造血的成熟血细胞谱系,而不诱导异常扩增(Yoshihara等人,2007; Spindler等人 ,2014; Hu等人,2016)。鹅卵石区域形成细胞试验(CAFC试验)是长期骨髓HSC和MSC的体外共培养试验。当培养MSC在组织培养皿中完成融合时,将HSC铺在MSC上(de Haan和Ploemacher,2002)。 CAFC测定与骨髓的体内研究相当,并且可用作快速筛选测定以测试HSC的干细胞活性和MSC的支持活性(Ploemacher等人, ...

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