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Sodium dodecyl sulfate

十二烷基硫酸钠

Company: Sigma-Aldrich
Catalog#: L3771
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Preparation of Cerebellum Granule Neurons from Mouse or Rat Pups and Evaluation of Clostridial Neurotoxin Activity and Their Inhibitors by Western Blot and Immunohistochemistry
Author:
Date:
2018-07-05
[Abstract]  Cerebellar Granule Neurons (CGN) from post-natal rodents have been widely used as a model to study neuronal development, physiology and pathology. CGN cultured in vitro maintain the same features displayed in vivo by mature cerebellar granule cells, including the development of a dense neuritic network, neuronal activity, neurotransmitter release and the expression of neuronal protein markers. Moreover, CGN represent a convenient model for the study of Clostridial Neurotoxins (CNT), most notably known as Tetanus and Botulinum neurotoxins, as they abundantly express both CNT receptors and intraneuronal substrates, i.e., Soluble N-ethylmaleimide-sensitive factor activating protein receptors (SNARE proteins). Here, we describe a protocol for obtaining a highly pure ... [摘要]  来自产后啮齿动物的小脑颗粒神经元(CGN)已被广泛用作研究神经元发育,生理学和病理学的模型。 CGN体外培养维持成熟小脑颗粒细胞在体内显示的相同特征,包括发育致密的神经炎网络,神经元活动,神经递质释放和神经元的表达 蛋白质标记。 此外,CGN代表了梭菌神经毒素(CNT)研究的便利模型,最着名的是破伤风和肉毒杆菌神经毒素,因为它们大量表达CNT受体和神经元内基质, ie ,可溶性N-乙基马来酰亚胺 - 敏感因子激活蛋白受体(SNARE蛋白)。 在这里,我们描述了从出生后大鼠/小鼠获得高纯度CGN培养物的方案和用CNT中毒的简便方法。 我们还说明了评估CNT活性及其抑制的方便方法。

【背景】梭菌神经毒素(CNT)的大家族由破伤风神经毒素(TeNT)和肉毒杆菌神经毒素(BoNT)的多种变体形成,它们分别是破伤风和肉毒中毒的神经麻痹毒素(Schiavo et al。,2000; Johnson和Montecucco,2008; Rossetto et al。,2014)。 TeNT,七种BoNT血清型(BoNT / A至/ G)及其许多亚型是金属蛋白酶,通过切割SNARE蛋白(可溶性N-乙基马来酰亚胺敏感因子激活蛋白受体),三种必需蛋白质来阻断神经递质的释放而引起神经麻痹。控制突触小泡与突触前质膜的融合(Rossetto et al。,2014; ...

ChIP-seq Experiment and Data Analysis in the Cyanobacterium Synechocystis sp. PCC 6803
Author:
Date:
2018-06-20
[Abstract]  Nitrogen is an essential nutrient for all living organisms. In cyanobacteria, a group of oxygenic photosynthetic bacteria, nitrogen homeostasis is maintained by an intricate regulatory network around the transcription factor NtcA. Although mechanisms controlling NtcA activity appear to be well understood, the sets of genes under its control (i.e., its regulon) remain poorly defined. In this protocol, we describe the procedure for chromatin immunoprecipitation using NtcA antibodies, followed by DNA sequencing analysis (ChIP-seq) during early acclimation to nitrogen starvation in the cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis). This protocol can be extended to analyze any DNA-binding protein in cyanobacteria for which suitable antibodies ... [摘要]  氮是所有生物体的必需营养素。 在蓝细菌中,一组含氧光合细菌通过围绕转录因子NtcA的错综复杂的调节网络维持氮稳态。 尽管控制NtcA活性的机制似乎已被很好地理解,但其控制下的基因集(即它的调节子)仍然没有很好的定义。 在该协议中,我们描述了使用NtcA抗体进行染色质免疫沉淀的过程,随后在蓝藻Synechocystis sp。早期适应氮饥饿期间进行DNA测序分析(ChIP-seq)。 PCC 6803(以下简称<集气囊)。 该协议可以扩展到分析蓝细菌中存在合适抗体的任何DNA结合蛋白。

【背景】为了维持体内平衡,细菌经常需要响应环境变化来调整基因表达。许多这些调整是由转录因子(TF)控制的,这些转录因子可以感知代谢信号并激活或抑制目标基因。然而,反映传统上费力的任务来表征TFs在体内的活性和范围,我们对它们在细菌中的结合位点的了解仍然有限。直到最近,染色质免疫沉淀与高通量测序分析的结合为快速确定基因组水平调节子打开了大门。特别是,ChIP-seq使用下一代测序(NGS)的能力来并行识别大量DNA序列。与微阵列相比,ChIP-seq的一个有吸引力的特征是对某些区域如启动子序列没有限制,并且可以研究整个基因组的TF结合位点。

在蓝细菌中,氮同化和代谢的全球调节剂是NtcA,属于CRP(cAMP受体蛋白)家族的TF(Herrero等人,2001)。在集胞蓝细菌中,NtcA通过将二聚体结合至包含共有序列GTAN ...

Total RNA Extraction from Dinoflagellate Symbiodinium Cells
Author:
Date:
2018-06-05
[Abstract]  Dinoflagellates are unicellular algae that can have photosynthetic or nonphotosynthetic lifestyles. Dinoflagellates in the genus Symbiodinium can enter endosymbiotic associations with corals, providing the metabolic basis for the highly productive and biologically diverse coral-reef ecosystems (Hoegh-Guldberg, 1999), as well as with other cnidarians, including sea anemones and jellyfish, and non-cnidarian hosts (Trench, 1993; Lobban et al., 2002; Mordret et al., 2016).

Here, I describe a protocol for isolating total RNA from Symbiodinium cells.
[摘要]  鞭毛藻是单细胞藻类,可以有光合或非光合生活方式。 Symbiodinium属中的甲藻类可以与珊瑚进入内共生关系,为高生产力和生物多样性珊瑚礁生态系统提供代谢基础(Hoegh-Guldberg,1999),以及其他的cnidarians,包括 海葵和海蜇,以及非刺猬寄主(Trench,1993; Lobban等人,2002; Mordret等人,2016)。

在这里,我描述了从Symbiodinium细胞中分离总RNA的方案。

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