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Ammonium hydroxide solution

氢氧化铵溶液

Company: Sigma-Aldrich
Catalog#: 320145
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Crude Preparation of Lipopolysaccharide from Helicobacter pylori for Silver Staining and Western Blot
Author:
Date:
2017-10-20
[Abstract]  This protocol provides an easy and rapid method to prepare lipopolysaccharide from the gastric pathogen Helicobacter pylori for visualization on acrylamide gels by silver staining and for detecting the presence of Lewis antigens by Western blot. The silver staining is a four-step procedure, involving a 20 min-oxidation step, a 10 min-silver staining step, a 2-10 min color development step and finally a 1-min color termination step. Lipopolysaccharide from H. pylori wild-type and corresponding mutants analyzed by this method are described in a recent publication (Li et al., 2017). This crude preparation of LPS for silver staining is also applicable in other Gram-negative bacteria. [摘要]  该方案提供了一种从胃病原幽门螺杆菌制备脂多糖的简便快速方法,用于通过银染显示丙烯酰胺凝胶,并通过Western印迹检测Lewis抗原的存在。 银染是四步法,包括20分钟氧化步骤,10分钟银染色步骤,2-10分钟显色步骤,最后是1分钟颜色终止步骤。 来自H的脂多糖。 在最近的出版物(Li等人,2017)中描述了通过该方法分析的野生型和相应的突变体的幽门螺杆菌。 这种用于银染的LPS的粗制剂也适用于其他革兰氏阴性细菌。
【背景】脂多糖(LPS)是一种大而可变的复合糖脂,其组成了大多数革兰氏阴性细菌外膜的外叶。 它通常由三个结构域组成:称为脂质A(或内毒素)的疏水结构域,其嵌入外膜中; 相对保守的非重复核 - 低聚糖; 和从细胞延伸到外部环境的可变O-抗原。 H的独特功能。 幽门螺杆菌脂多糖O抗原是模拟人Lewis抗原的岩藻糖基化寡糖结构的存在。 从革兰氏阴性细菌大规模提取高纯度的LPS是劳动密集型和耗时的。 在这里,在这个协议中,我们详细地描述了使用来自胃病原幽门螺杆菌的容易和快速的粗制备制剂用于通过银染和Lewis抗原通过蛋白质印迹进行表达。

Determination of VPS34/PIK3C3 Activity in vitro Utilising 32P-γATP
Author:
Date:
2016-08-20
[Abstract]  VPS34 is the only class III phosphatidylinositol-3-kinase (PI3K) in mammalian cells and produces the vast majority of cellular phosphatidylinositol-3-phosphate [PI(3)P]. PI(3)P is a key signalling lipid that plays many membrane trafficking roles in processes such as endocytosis and autophagy. VPS34 is a key cellular regulator, loss of function can have catastrophic effects and is embryonic lethal (Zhou et al., 2011). The levels of cellular PI(3)P can be determined by fluorescent staining techniques and can be used to monitor effects upon VPS34 activity, however it is important to verify that any changes are mediated by VPS34, particularly as alternate pathways of PI(3)P production are possible such as via class II PI3Ks (Devereaux et al., 2013). Assaying VPS34 activity ... [摘要]  VPS34是哺乳动物细胞中唯一的III类磷脂酰肌醇-3-激酶(PI3K),并且产生绝大多数细胞磷脂酰肌醇-3-磷酸[PI(3)P]。 PI(3)P是一个关键的信号脂质,在诸如内吞作用和自噬的过程中起许多膜运输的作用。 VPS34是关键的细胞调节剂,功能丧失可具有灾难性作用并且是胚胎致死的(Zhou等人,2011)。 细胞PI(3)P的水平可以通过荧光染色技术确定,并且可以用于监测对VPS34活性的影响,然而重要的是验证任何变化由VPS34介导,特别是作为PI(3)的替代途径, P生产是可能的,例如通过II类PI3K(Devereaux等人,2013)。 直接在体外测定VPS34活性可以是描绘特定刺激的作用的关键阶段。

A Phosphopeptide Purification Protocol for the Moss Physcomitrella paten
Author:
Date:
2015-07-20
[Abstract]  Protein phosphorylation is one of the most common post-translational modifications in eukaryotic cells and plays a critical role in a vast array of cellular processes. Efficient methods of protein extraction and phosphopeptide purification are required to ensure the detection of high quality of proteins. In our hands, phenol extraction of proteins and TiO2 chromatography enrich phosphorylated peptides more efficiently than other methods in the moss Physcomitrlla patens (P. patens). [摘要]  蛋白磷酸化是真核细胞中最常见的翻译后修饰之一,并在大量细胞过程中起关键作用。 需要蛋白质提取和磷酸肽纯化的有效方法以确保高质量的蛋白质的检测。 在我们的手中,蛋白质和TiO 2层析的酚提取比在小麦Physcomitrlla patens中的其它方法更有效地富集磷酸化肽。 )。

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