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Na2HPO4·7H2O

磷酸氢二钠

Company: Sigma-Aldrich
Catalog#: S7907
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Identification of Socially-activated Neurons
Author:
Date:
2020-09-05
[Abstract]  Determining the neuronal circuitry responsible for specific behaviors is a major focus in the field of neurobiology. Activity-dependent immediate early genes (IEGs), transcribed and translated shortly after neurons discharge action potentials, have been used extensively to either identify or gain genetic access to neurons and brain regions involved in such behaviors. By using immunohistochemistry for the protein product of the IEG c-Fos combined with retrograde labeling of specific neuronal populations, precise experimental timing, and identical data acquisition and processing, we present a method to quantitatively identify specific neuronal subpopulations that were active during social encounters. We have previously used this method to show a stronger recruitment of ventral hippocampal ... [摘要]  [摘要] 确定负责特定行为的神经元回路是神经生物学领域的主要重点。在神经元释放动作电位后不久就转录和翻译的依赖活动的立即早期基因(IEG)已被广泛用于识别或获得涉及此类行为的神经元和大脑区域的遗传途径。通过对IEG c- Fos 的蛋白质产物使用免疫组织化学,结合特定神经元群体的逆向标记,精确的实验时间,相同的数据采集和处理 ng,我们提出了一种方法,可以定量地识别在社交活动中活跃的特定神经元亚群。与社交活动后投射到下丘脑外侧的神经海马神经元相比,我们以前曾使用这种方法显示出投射到内侧前额叶皮层的腹侧海马神经元更强的募集。在优化用于逆行示踪剂注射的手术后,该方法将有助于识别和定位参与许多不同行为的神经元群体。

[背景] 神经生物学领域的基本目标是确定大脑如何处理感觉输入并产生行为输出命令。在过去的二十年中,某些大脑区域已成为控制特定行为的典范实例,例如海马体用于位置记忆(Bird和Burgess,2008年)。最近的工作开始以可以推广到多种行为的方式来解析这些区域的特定功能(Aronov 等人,2017),并确定在由远程神经元连接的给定行为中多个大脑区域的作用投影( ...

High-level Production of Recombinant Membrane Proteins Using the Engineered Escherichia coli Strains SuptoxD and SuptoxR
Author:
Date:
2020-08-05
[Abstract]  We have previously described the development of two specialized Escherichia coli strains for high-level recombinant membrane protein (MP) production. These engineered strains, termed SuptoxD and SuptoxR, are capable of suppressing the cytotoxicity caused by MP overexpression and of producing greatly enhanced MP yields. Here, we present a Bio-protocol that describes gene overexpression and culturing conditions that maximize the accumulation of membrane-integrated and well-folded recombinant MPs in these strains. [摘要]  [摘要] 我们之前已经描述了两种用于生产高水平重组膜蛋白(MP)的大肠杆菌菌株的开发。这些工程菌株,称为SuptoxD和SuptoxR,能够抑制MP过度表达引起的细胞毒性,并产生显著提高的MP产量。在这里,我们提出一个生物协议,描述基因过度表达和培养条件,最大限度地积累膜整合和折叠良好的重组多磺酸粘多糖在这些菌株。

[背景]多磺酸粘多糖在所有活生物体的细胞中执行多种关键功能(Wagner et al.,2006;Schlegel et al.,2010),是当前和未来药物的主要靶点(Yildrim et al.,2007)。获得足够数量的分离蛋白是进行生化和结构研究的前提,这反过来又可以加深对其功能的理解,并发现新的MP靶向药物。

由于多磺酸粘多糖通常在其天然环境中以极低的丰度出现,异源宿主通常用于其重组过表达和随后的纯化。许多不同的系统已被用作原核和真核来源的多种多磺酸粘多糖的过表达宿主(Wagner等人,2006年)。其中,大肠杆菌是最受欢迎的一种,因为它的成本非常低,使用方便(Makino等人,2011年)。事实上,这种细菌已经成功地用于生产储存在蛋白质数据库中的所有重组产生的MP结构的大约20%(Dilworth等人,2018年)。尽管有这些优势和成功,但使用大肠杆菌作为MP生产的异源宿主通常伴随着严重的毒性、低水平的最终生物量和微小的最终产量(Miroux和Walker,1996;Wagner等人,2007;Link等人,2008;Gubellini等人,2011)。 ...

Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae
Author:
Date:
2020-03-05
[Abstract]  Superoxide dismutases (SODs) act as a primary defence against reactive oxygen species (ROS) by converting superoxide anion radicals (O2-) into molecular oxygen (O2) and hydrogen peroxide (H2O2). Members of this enzyme family include CuZnSODs, MnSODs, FeSODs, and NiSODs, depending on the nature of the cofactor that is required for proper activity. Most eukaryotes, including yeast, possess CuZnSOD and MnSOD. This protocol aims at assessing the activity of the yeast Saccharomyces cerevisiae MnSOD Sod2p from cellular extracts using nitroblue tetrazolium staining. This method can be used to estimate the cellular bioavailability of Mn2+ as well as to evaluate the redox state of the cell. [摘要]  [摘要 ] 超氧化物歧化酶(SOD能)充当主防御针对反应性氧物质(ROS)通过转换的超氧阴离子自由基(O 2 - )为分子氧(O 2 )和过氧化氢(H 2 ? 2 )。这种酶的家庭成员包括CuZnSODs ,MnSODs ,FeSODs 和NiSODs ,这取决于是需要适当的活动辅助因子的性质。大多数真核生物,包括酵母,都具有CuZnSOD 和MnSOD 。该协议旨在评估酵母的活性 使用硝基蓝四唑染色法从细胞提取物中提取酿酒酵母MnSOD Sod2p 。该方法可用于估计Mn 2+ 的细胞生物利用度以及评估细胞的氧化还原状态。

[背景 ] 的SODs被定义为减少正常有氧代谢为氧气和过氧化氢期间形成的氧的有害自由基含金属的抗氧化剂酶。:这些酶是基于需要作为辅因子进行适当的酶活性的金属分类CuZnSODs ,MnSODs ,FeSODs ,和NiSODs 。在酿酒酵母中,有两个S OD :CuZn-Sod1p和Mn-Sod2p(Abreu和Cabelli ...

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