{{'Search' | translate}}
 

Falcon® 15 mL Polystyrene Centrifuge Tube, Conical Bottom, with Dome Seal Screw Cap, Sterile

Falcon ® 15 mL聚苯乙烯离心管,圆锥底,带圆顶密封螺帽

Company: Corning
Catalog#: 352095
Bio-protocol()
Company-protocol()
Other protocol()

Chromatin Immunoprecipitation Experiments from Whole Drosophila Embryos or Larval Imaginal Discs
Author:
Date:
2017-06-05
[Abstract]  Chromatin Immunoprecipitation coupled either to qPCR (qChIP) or high-throughput sequencing (ChIP-Seq) has been extensively used in the last decades to identify the DNA binding sites of transcription factors or the localization of various histone marks along the genome. The ChIP experiment generally includes 7 steps: collection of biological samples (A), cross-linking proteins to DNA (B), chromatin isolation and fragmentation by sonication (C), sonication test (D), immunoprecipitation with antibodies against the protein or the histone mark of interest (E), DNA recovery (E), identification of factor-associated DNA sequences by PCR or sequencing (F). The protocol described here can readily be used for ChIP-seq and ChIP-qPCR experiments. The entire procedure, describing experimental setup ... [摘要]  与qPCR(qChIP)或高通量测序(ChIP-Seq)相结合的染色质免疫沉淀已被广泛用于识别转录因子的DNA结合位点或基因组中各种组蛋白标记的定位。 ChIP实验通常包括7个步骤:收集生物样品(A),交联蛋白质到DNA(B),染色质分离和通过超声处理分离(C),超声处理测试(D),用针对蛋白质的抗体进行免疫沉淀感兴趣的组蛋白标记(E),DNA回收(E),通过PCR或测序鉴定因子相关DNA序列(F)。这里描述的协议可以容易地用于ChIP-seq和ChIP-qPCR实验。描述在完整的果蝇组织中优化分析的实验设置条件的整个过程可以在四天内完成。

背景 尽管永生化的培养细胞广泛用于研究各种细胞类型的染色质景观,但是在生理条件下在体内探测相互作用的有价值的方法对于进行转录的时间或空间比较分析是必要的因子和组蛋白修饰图在不同阶段的果蝇发展或不同组织之间。在这里,我们提供了一个详细的ChIP协议,已被优化,以便在整个果蝇胚胎和幼虫成像光盘上工作,突出关键的实验参数。

Measuring Caenorhabditis elegans Sleep during the Transition to Adulthood Using a Microfluidics-based System
Author:
Date:
2017-03-20
[Abstract]  C. elegans sleep during development is regulated by genes and cellular mechanisms that are conserved across the animal kingdom (Singh et al., 2014; Trojanowski and Raizen, 2016). C. elegans developmental sleep is usually assessed during the transition to adulthood, a 2.6 h time interval called lethargus (Raizen et al., 2008; Singh et al., 2011). During lethargus, animals cycle between periods of immobility (sleep bouts) and periods of active locomotion (motion bouts). Sleep bouts resemble sleep in other species based on behavioral criteria, including cessation of feeding and locomotion, increased arousal threshold for response to sensory stimulation, rapid reversibility, and homeostatic response to sleep loss. Several assays have been developed ... [摘要]  C。发展过程中的线虫睡眠受到动物界保护的基因和细胞机制的限制(Singh等人,2014; Trojanowski和Raizen,2016)。 C。线虫发育睡眠通常在成年过渡期间进行评估,2.6h时间间隔称为lethargus(Raizen等人,2008; Singh等人, 2011)。在嗜睡期间,动物在不动的时期(睡眠开始)和积极运动时期(运动发作)之间循环。睡眠状态基于行为标准,包括停止进食和运动,增加唤醒阈值以响应感觉刺激,快速可逆性和对睡眠损失的稳​​态反应,类似于其他物种的睡眠。已经开发了几种用于在C中研究睡眠的测定法。 (Belfer等人,2013; Bringmann,2011; Nelson等人,2013; Raizen等人, 2008)。在这里,我们提供了一个详细的评估方案。线虫在lethargus期间睡眠,许多研究组已经成功使用,结合简单的微流体室,具有照明系统的低成本照相机和基于图像减法的计算分析。我们注意到,这个系统可以很容易地适应于评估任何小动物的睡眠。

背景 C。睡眠通常根据运动停止进行评估,这是整个动物界睡眠的常见特征。由于C期间睡眠发作的间歇性。线虫发育睡眠,计算机视觉通常用于跟踪C的活动。线虫在lethargus期间。动物被约束到单个焦平面以保持它们的焦点。因为C.线虫可以通过长时间的游泳用尽液体(Ghosh和Emmons,2008),C。线虫 ...

An Acute Mouse Spinal Cord Slice Preparation for Studying Glial Activation ex vivo
Author:
Date:
2017-01-20
[Abstract]  Pathological conditions such as amyotrophic lateral sclerosis, spinal cord injury and chronic pain are characterized by activation of astrocytes and microglia in spinal cord and have been modeled in rodents. In vivo imaging at cellular level in these animal models is limited due to the spinal cord’s highly myelinated funiculi. The preparation of acute slices may offer an alternative and valuable strategy to collect structural and functional information in vitro from dorsal, lateral and ventral regions of spinal cord. Here, we describe a procedure for preparing acute slices from mouse spinal cord (Garré et al., 2016). This preparation should allow further understanding of how glial cells in spinal cord respond acutely to various inflammatory challenges. [摘要]  病理状况如肌萎缩性侧索硬化,脊髓损伤和慢性疼痛的特征在于脊髓中星形胶质细胞和小胶质细胞的活化,并已在啮齿动物中建模。 在这些动物模型中的细胞水平的体内成像由于脊髓的高度髓鞘化的功能而受到限制。 急性切片的制备可能提供一种替代和有价值的策略,从脊髓的背侧,外侧和腹侧区域体外收集结构和功能信息。 在这里,我们描述了一种从小鼠脊髓制备急性切片的过程(Garréet al。,2016)。 这个准备应该进一步了解脊髓中的神经胶质细胞是如何急剧地反应各种炎症的挑战。
【背景】已经使用小鼠转基因技术来模拟影响脊髓的不同人类病态,其中许多特征为局部胶质激活,神经炎症的一个标志。利用基于激光扫描显微技术的共焦显微镜(White et al。,1987)和双光子显微镜(Denk等,1990),大大提高了对健康和疾病中胶质生物学的认识。 )以无创的方式可视化活体动物中的细胞结构和亚细胞结构域;例如,表达遗传编码的记者或钙传感器的小鼠已被用于分别形成胶质结构(体细胞和过程)并研究钙动力学和信号传导(Davalos等人,2005; ...

Comments