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Eagle's Minimum Essential Medium

低限量 Eagle 培养基

公司名称: ATCC
产品编号: 30-2003
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Adhesion and Invasion Assay Procedure Using Caco-2 Cells for Listeria monocytogenes
Author:
Date:
2017-05-05
[Abstract]  Listeria monocytogenes is an important Gram-positive foodborne pathogen that is a particular problem in ready-to-eat food. It has an ability to survive in harsh conditions like refrigeration temperatures and high salt concentrations and is known to cross intestinal, placental and blood-brain barriers. Several cancerous cell lines like cervical, liver, dendritic, intestinal and macrophages have been used to study in vitro propagation and survival of listeria in human cells. Human intestinal epithelial cells have been used to study how listeria crosses the intestinal barrier ... [摘要]  单核细胞增生利斯特氏菌是一种重要的革兰氏阳性食源性病原体,是即食食品中的一个特殊问题。它具有在诸如制冷温度和高盐浓度的恶劣条件下生存的能力,并且已知可以穿过肠,胎盘和血脑屏障。已经使用了诸如子宫颈,肝脏,树突状细胞,肠和巨噬细胞的几种癌细胞系来研究人细胞中李斯特菌的体外扩增和存活。人肠上皮细胞已被用于研究李斯特菌如何穿过肠屏障并引起感染。本文中的方案描述了生长Caco-2细胞的过程,维持细胞并将其用于粘附和侵袭测定。在粘附测定期间,将细胞与李斯特菌孵育30分钟,但是在侵袭测定中,细胞生长在感染后的几个时间点被停止以监测细胞中李斯特菌的生长和存活率。

背景 ...

Fluorescence Microscopy Analysis of Drug Effect on Autophagosome Formation
Author:
Date:
2014-04-05
[Abstract]  The autophagy protein, LC3 represents a reliable characteristic marker for autophagosomal structures. The initial LC3 is processed by the cysteine protease autophagy-related gene 4 (Atg4) at its C terminus in order to create LC3-I generally localized in the cytoplasm. Afterwards LC3-I is conjugated with phosphatidylethanolamine (PE) to become LC3-PE or LC3-II predominantly localised on the autophagosomal membranes (outer and inner). Autolysosomal content of LC3-II is very low as upon autophago/lysosomal fusion it is either cleaved off from the outer membrane by Atg4 or degraded together with ... [摘要]  自噬蛋白,LC3代表自噬体结构的可靠特征标记。最初的LC3由半胱氨酸蛋白酶自噬相关基因4(Atg4)在其C末端处理,以产生通常位于细胞质中的LC3-I。之后LC3-I与磷脂酰乙醇胺(PE)缀合以变成主要定位在自噬体膜(外部和内部)上的LC3-PE或LC3-II。 LC3-II的自溶酶体内含物非常低,因为在自噬/溶酶体融合时,其被Atg4从外膜切割或与内膜一起通过溶酶体活性降解。因此,GFP-LC3和mCherry-GFP-LC3可能通过常规或共聚焦荧光显微镜(FM)可视化。在这种情况下,mCherry-GFP-LC3或GFP-LC3细胞质池被可视化为均匀分散的信号,并且含有mCherry-GFP-LC3-II或GFP-LC3-II的自噬体被检测为点状形成。斑点数可以用作自噬体丰度的标志物。一般来说,我们建议计数每个细胞的GFP-LC3斑点的平均数。

Flow Cytometric Analysis of Autophagic Activity with Cyto-ID Staining in Primary Cells
Author:
Date:
2014-04-05
[Abstract]  Flow cytometry allows very sensitive and reliable high-throughput analysis of autophagic flux. This methodology permits to screen cells in flow and capture multi-component images. Using this technology autophagic flux may be analysed accurately in both suspension as well as adherent cells upon trypsinization independent of how heterogeneous the autophagosomal content might be. The method is based on Cyto-ID staining of autophagic compartments (pre-autophagosomes, autophagosomes, and autophagolysosomes) in live cells using Cyto-ID® Autophagy Detection Kit. Autophagic compartments ... [摘要]  流式细胞术允许非常灵敏和可靠的高通量分析自噬通量。该方法允许在流动中筛选细胞并捕获多组分图像。使用这种技术,可以在悬浮液以及粘附细胞中在胰蛋白酶消化时精确分析自噬流,而与自噬体内容物的异质性无关。该方法基于使用Cyto-ID自动吞噬检测试剂盒的活细胞中的自体吞噬细胞(前自噬体,自噬体和自噬溶酶体)的Cyto-ID染色。自噬区室是动态溶酶体降解过程的中间组分,并且它们在特定时间点的细胞内丰度是其产生和降解之间建立的平衡的函数。自噬吞噬的确定有助于自噬体形成的早期诱导和自噬体成熟的晚期抑制之间的区分,因为两者都导致自噬体存在的最终增加。 Cyto-ID测定基于使用选择性染色自噬隔室的特定染料,因此允许测定自噬通量,作为染色隔室在碱性或活化条件[雷帕霉素(1-5μmol/L),PP242(1- (NH 4 Cl)(10-20μmol/L)或含有6mmol/L葡萄糖(饥饿培养基)的Hanks平衡盐溶液),使用溶酶体化合物阻断自噬溶酶体降解mmol/L)或氯喹(CQ)(5-10μmol/L)。 ΔMFICyto-ID = MFI Cyto-ID(+ CQ/NH 4 Cl)-MFI Cyto-ID(-CQ/NH 4 Cl)。

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