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PierceTM BCA Protein Assay Kit

Pierce TM BCA蛋白测定试剂盒

Company: Thermo Fisher Scientific
Catalog#: 23225
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Analysis of Metals in Whole Cells, Thylakoids and Photosynthetic Protein Complexes in Synechocystis sp. PCC6803
Author:
Date:
2018-06-20
[Abstract]  Metals are essential in many biological processes, including oxygenic photosynthesis. Here we described a method to measure the metal pool in whole cells and thylakoids, including the bioactive pool in intact photosynthetic protein complexes in the model oxygenic cyanobacterium Synechocystis PCC6803. In the first part of the protocol, whole cells and thylakoid membranes are carefully prepared, in which the total metal concentrations are measured by inductively coupled plasma triple-quadrupole mass spectrometry (ICP-QQQ-MS). In the second part of the protocol, isolated thylakoids are solubilized to release the integral membrane proteins and the metal binding protein complexes. These intact photosynthetic protein complexes are subjected to size exclusion chromatography (SEC) and ... [摘要]  金属在许多生物过程中是必不可少的,包括含氧光合作用。 在这里我们描述了测量全细胞和类囊体中金属库的方法,包括模型含氧蓝藻PCH6803中完整光合蛋白复合体中的生物活性库。 在方案的第一部分中,仔细制备全细胞和类囊体膜,其中通过电感耦合等离子体三重四极杆质谱(ICP-MS / MS)测量总金属浓度。 在该方案的第二部分,分离的类囊体被溶解以释放完整的膜蛋白和金属结合蛋白复合物。 将这些完整的光合蛋白复合物进行尺寸排阻色谱(SEC),并通过ICP-MS / MS进行连接分析尺寸分离的复合物中的金属结合。

【背景】氧光合作用的过程需要金属,因为它们在光合电子传递链中作为辅因子和催化剂的基本功能。光合装置需要Fe-S簇,血红素桥Fe和非血红素Fe形式的铁(Fe),可溶性移动电子载体蛋白质质体蓝素中的铜(Cu),叶绿素中的镁(Mg), ...

In vitro Analysis of Ubiquitin-like Protein Modification in Archaea
Author:
Date:
2018-05-20
[Abstract]  The ubiquitin-like (Ubl) protein is widely distributed in Archaea and involved in many cellular pathways. A well-established method to reconstitute archaeal Ubl protein conjugation in vitro is important to better understand the process of archaeal Ubl protein modification. This protocol describes the in vitro reconstitution of Ubl protein modification and following analysis of this modification in Haloferax volcanii, a halophilic archaeon serving as the model organism. [摘要]  泛素样(Ubl)蛋白广泛分布于古细菌中并参与许多细胞途径。 为了更好地理解古细菌Ub1蛋白质修饰的过程,重建体外古细菌Ubl蛋白质缀合物的完善方法是很重要的。 该协议描述了Ubl蛋白质修饰的体外重建以及在作为模型生物的嗜盐古细菌Haloferax volcanii 中对这种修饰进行分析。

【背景】泛素(Ub)与靶蛋白共价连接的过程被称为泛素化,其控制真核细胞中大量的细胞过程(Glickman和Ciechanover,2002; Komander和Rape,2012)。遍在蛋白化由一系列酶(包括Ub激活酶(E1),Ub结合酶(E2s)和Ub连接酶(E3s))催化。泛素化的体外重建是确定酶之间或E3与蛋白质底物之间特异性的有用测定法(Zhao等人,2012)。在古细菌中,Ubl蛋白SAMP采用Ub折叠,并且与E1样酶UbaA催化的蛋白靶标异肽连接[Maupin-Furlow,(2014)综述]。尽管E1同系物在古细菌中广泛存在,但基于一级序列比较,在大多数古细菌中未预测经典E2或E3酶。我们最近对Haloferax volcanii的研究表明甲硫氨酸亚砜还原酶A(MsrA)是Ubl蛋白质修饰(sampylation)与UbaA一起在体内温和的氧化条件下和< (体外)(fu="">

Superoxide Dismutase (SOD) and Catalase (CAT) Activity Assay Protocols for Caenorhabditis elegans
Author:
Date:
2017-08-20
[Abstract]  Assays for superoxide dismutase (SOD) and catalase (CAT) activities are widely employed to indicate antioxidant responses underlying the toxic effects of test chemicals. Yet, earlier studies mainly described the procedures as performed according to manufacturer’s instructions without modifications that are specific to any organisms. The present protocol describes the steps in analyzing the superoxide dismutase (SOD) and catalase (CAT) activities in C. elegans, which is a model organism that can be used to study effects of pharmaceutical compounds and environmental pollutants. The main steps include: (1) sample preparation; (2) total protein assay; (3) SOD activity assay; (4) CAT activity assay; and (5) medium list and formula, and also data analysis and performance notes. [摘要]  超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性的测定被广泛用于表明测试化学品的毒性作用的抗氧化反应。 然而,早期的研究主要描述了根据制造商的说明进行的程序,而无需对任何生物体特异的修改。 本方案描述了分析线虫中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性的步骤,它是可用于研究药物化合物和环境污染物的影响的模型生物。 主要步骤包括:(1)样品制备; (2)总蛋白测定; (3)SOD活性测定; (4)CAT活性测定; 和(5)中等名单和公式,以及数据分析和绩效说明。
【背景】生物标志物是对化学,药剂或治疗干预的响应而检查生物和病原过程至关重要的。生物体内的各种生物过程导致引起氧化应激的活性氧(ROS)。为了应对这种氧化应激,生物体可以部署超氧化物歧化酶(SOD)和过氧化氢酶(CAT)以清除ROS,以保护细胞的稳态(Balaban等,2005)。一方面,各种化学物质(污染物)可以阻止这种抗氧化反应,并扰乱包括人类在内的生物体的健康。另一方面,许多药物旨在加强抗氧化反应以改善健康。因此,SOD和CAT的活动对于反映化学品或/和药物的潜在影响非常重要。
   秀丽隐杆线虫(秀丽隐杆线虫)是一种用于研究药物化合物(Dengg和van Meel,2004; Carretero等,2017)和环境污染物(Yu et ...

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