{{'Search' | translate}}
 

Stericup Quick Release-GP Sterile Vacuum Filtration System

公司名称: Merck
产品编号: S2GPU11RE
Bio-protocol()
Company-protocol()
Other protocol()

Expression and Purification of Recombinant Skd3 (Human ClpB) Protein and Tobacco Etch Virus (TEV) Protease from Escherichia coli
Author:
Date:
2020-12-05
[Abstract]  

Skd3 (encoded by human CLPB) is a mitochondrial AAA+ protein comprised of an N-terminal ankyrin-repeat domain and a C-terminal HCLR-clade nucleotide-binding domain. The function of Skd3 has long remained unknown due to challenges in purifying the protein to high quality and near homogeneity. Recently we described Skd3 as a human mitochondrial protein disaggregase that solubilizes proteins in the mitochondrial intermembrane space. This protocol overcomes the challenges associated with purifying Skd3 and allows for in depth in vitro study of Skd3 activity. Tobacco etch virus (TEV) protease is

...
[摘要]  [摘要] Skd3(由人类CLPB编码)是一种线粒体AAA +蛋白,由N末端锚蛋白重复域和C末端HCLR分支核苷酸结合域组成。由于在纯化蛋白质达到高质量和接近均质性方面的挑战,Skd3的功能长期未知。最近,我们描述Skd3作为人类线粒体蛋白disaggregase ,在线粒体膜间间隙增溶蛋白。该协议克服了与纯化Skd3相关的挑战,并允许对Skd3活性进行深入的体外研究。Skd3的纯化需要烟草蚀刻病毒(TEV)蛋白酶。因此,我们还描述了如何净化高质量TEV蛋白酶可用于纯化Skd3,其他纯化方案以及需要TEV蛋白酶的体外测定。


[背景] Skd3是一种线粒体AAA +蛋白,与多系统线粒体疾病VII型3-甲基谷氨酸酸尿症(MGCA7)有关(Capo-Chichi等人,2015; Kanabus等人,2015; Saunders等人,, 2015; Wortmann等人,2015 ; Kiykim等人,2016 )。由于在体外研究Skd3的能力有限,因此对生物学功能和这些突变对Skd3活性的影响的研究仍难以捉摸(Cupo和Shorter,2020; ...

Giant Mimiviridae CsCl Purification Protocol
Author:
Date:
2020-11-20
[Abstract]  While different giant viruses’ purification protocols are available, they are not fully described and they use sucrose gradient that does not reach an equilibrium. Here, we report a protocol for the purification of members of the Mimiviridae family virions resulting from Acanthamoeaba castellanii infections. Viruses are harvested after cell lysis and purified through a high density CsCl gradient to optimize the isolation of the virus from the cell debris or other potential contaminants. Due to the large size of the virion capsids, reaching half a micrometer diameter, ... [摘要]  [摘要]虽然有不同的巨型病毒纯化方案,但它们并没有被完全描述,它们使用的蔗糖梯度不能达到平衡。在这里,我们报告了一个方案,用于纯化由棘球绦虫感染引起的拟病毒科成员病毒。病毒在细胞裂解后收获,并通过高密度CsCl梯度进行纯化,以优化从细胞碎片或其他潜在污染物中分离病毒。由于病毒粒子衣壳尺寸大,直径可达半微米,因此可以通过光学显微镜来监控工艺的质量。由此得到的纯化颗粒可用于进行新的感染、DNA提取、结构研究、糖成分分析、亚组分鉴定或蛋白质组学实验。

[背景]Mimivirus是第一种在光学显微镜下可见的病毒,在大小和基因组复杂度上与单细胞生物重叠,这一发现开创了病毒学的新研究领域(La Scola et al.,2003;Raoult et al.,2004)。在过去的15年里,许多额外的米病毒科成员被从各种环境中分离出来,并且已经发布了几种纯化病毒离子的方案(La Scola等人,2003年;Byrne等人,2009年;Arslan等人,2011年;Philippe等人,2013年;Campos等人,2014年;Andrade等人,2017年)。开发了不同的方法,主要涉及蔗糖缓冲(Campos et al.,2014;Andrade et al.,2017)或蔗糖不连续梯度(Arslan et ...

产品评论