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DAKO fluoresence mounting media

Company: Dako
Catalog#: S302380-2
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Ciliary Assembly/Disassembly Assay in Non-transformed Cell Lines
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Date:
2018-03-20
[Abstract]  The primary cilium is a non-motile sensory organelle whose assembly and disassembly are closely associated with cell cycle progression. The primary cilium is elongated from the basal body in quiescent cells and is resorbed as the cells re-enter the cell cycle. Dysregulation of ciliary dynamics has been linked with ciliopathies and other human diseases. The in vitro serum-stimulated ciliary assembly/disassembly assay has gained popularity in addressing the functions of the protein-of-interest in ciliary dynamics. Here, we describe a well-tested protocol for transfecting human retinal pigment epithelial cells (RPE-1) and performing ciliary assembly/disassembly assays on the transfected cells. [摘要]  主要纤毛是一种非运动感觉细胞器,其装配和拆卸与细胞周期进程密切相关。 初级纤毛在静止细胞中从基体拉长并随着细胞重新进入细胞周期而被吸收。 睫状动力失调与纤毛病和其他人类疾病有关。 体外血清刺激的睫状体装配/分解测定已经在解决睫状动力学中感兴趣的蛋白质的功能方面受到欢迎。 在这里,我们描述了转染人视网膜色素上皮细胞(RPE-1)和对转染细胞进行睫状体装配/分解测定的充分测试的方案。

【背景】初级纤毛是毛发样感觉细胞器,其在G 0 / G 1期出现,并且在细胞周期的S期之前分解(Tucker等, et al。,1979)。先前的研究已经证实,某些未转化的细胞类型(即,甚至是RPE-1细胞,3T3成纤维细胞和小鼠胚胎成纤维细胞[MEFs])可以被饿死以诱导静止和睫状体形成。随后的血清再次添加触发双相睫状体吸收,其在刺激后2小时和24小时达到峰值(Tucker等人,1979; Li等人,2011) 。该现象为文献中常用的血清刺激的睫状体组装/分解测定奠定了基础,以鉴定参与睫状体组装和拆卸的蛋白质(Pugacheva等人,2007; ...

Mouse Model of Immune Complex-mediated Vasculitis in Dorsal Skin and Assessment of the Neutrophil-mediated Tissue Damage
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Date:
2017-12-20
[Abstract]  Neutrophils are the most abundant leukocytes in the blood. In the recent decades, their crucial roles in host defense, immune regulation and tissue damage have been studied in a deeper dimension. In this protocol, we described a mouse model of immune complex-mediated vasculitis in the dorsal skin induced by Arthus reaction, and the subsequent analysis of edema, hemorrhage and tissue damage due to neutrophil activation by means of Evans blue area analysis, histology, and immunofluorescence. This protocol could facilitate the investigation of cellular therapy strategy against over-activated neutrophil-mediated tissue damage. [摘要]  嗜中性粒细胞是血液中最丰富的白细胞。 近几十年来,它们在宿主防御,免疫调节和组织损伤方面的关键作用已经得到了更深入的研究。 在这个协议中,我们描述了Arthus反应诱导的背部皮肤中的免疫复合物介导的血管炎的小鼠模型,以及随后通过伊文思蓝区域分析,组织学和分析由于嗜中性粒细胞活化引起的水肿,出血和组织损伤免疫荧光。 该协议可以促进细胞治疗策略对过度活化的嗜中性粒细胞介导的组织损伤的调查。

【背景】嗜中性粒细胞构成循环白细胞的最大的进化保守部分。他们引导第一波主机防御感染或组织损伤。嗜中性粒细胞介导的细胞毒性的体外模型已被充分证实(Incani等人,1981; Dallegri等人,1984; Saffarzadeh等人,等人,2012年)。但是,为了剖析中性粒细胞介导的无菌组织损伤的复杂性,“体内”模型是必不可少的。

免疫复合物(IC)介导的血管炎是由抗原 - 抗体复合物在血管中沉积引发的疾病,其随后导致补体激活,嗜中性粒细胞募集和活化。活化的嗜中性粒细胞释放的大量活性氧和蛋白酶损害血管壁的内皮衬里并导致水肿和出血(Sindrilaru等人,2007; Goerge等人, ,2008; ...

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