{{'Search' | translate}}
 

Laser scanner

Company: Cytiva
Catalog#: Typhoon FLA 9500
Bio-protocol()
Company-protocol()
Other protocol()

Relative Quantification of NaV1.1 Protein in Mouse Brains Using a Meso Scale Discovery-Electrochemiluminescence (MSD-ECL) Method
Author:
Date:
2021-02-05
[Abstract]  

Densitometric analysis is often used to quantify NaV1.1 protein on immunoblots, although the sensitivity and dilution linearity of the method are usually poor. Here we present a protocol for quantification of NaV1.1 in mouse brain tissues using a Meso Scale Discovery-Electrochemiluminescence (MSD-ECL) method. MSD-ECL is based on ELISA (enzyme-linked immunosorbent assay) and uses electrochemiluminescence to produce measurable signals. Two different antibodies are used in this assay to capture and detect NaV1.1 respectively in brain tissue lysate. The specificity of the antibodies is confirmed by Scn1a gene knock-out tissue. The calibration curve standards used in this assay were generated with mouse liver lysate spiked with mouse brain lysate, instead of using a recombinant protein. We

...
[摘要]  [摘要]尽管该方法的灵敏度和稀释度线性通常较差,但经常使用光密度分析法定量免疫印迹上的Na V 1.1蛋白。这里瓦特E存在的Na进行定量的协议V在使用小鼠脑组织1.1细观量表发现-电化学发光(MSD-ECL)方法。MSD-ECL基于ELISA(酶联免疫吸附测定),并使用电化学发光产生可测量的信号。此测定法中使用了两种不同的抗体来捕获和检测Na V 1.1分别在脑组织中溶解。Scn1a基因敲除组织证实了抗体的特异性。此测定法中使用的校准曲线标准品是用掺有小鼠脑裂解液的小鼠肝裂解液而不是重组蛋白生成的。我们证明该方法是合格的,可用于特异性,准确度和精密度定量的小鼠脑组织中的Na V 1.1 。


[背景]的Na V 1.1,也称为电压门控钠通道的α亚基,I型,是一种跨膜通过对编码蛋白SCN1A基因(Meisler等人,2010 )。功能性Na V 1.1的表达降低会导致Dravet综合征(DS),这是一种严重的早发性癫痫性脑病(Dravet等,2005)。Na V 1.1在生物样品中的表达已用作DS的非临床药理生物标记物,可以使用免疫印迹的光密度分析进行测量。密度测定法通常不如标准免疫测定法准确和灵敏。此外,由于蛋白质序列的同源性,某些Na V 1.1抗体可能会与其他电压门控钠通道(VGSC)发生交叉反应,包括Na V 1.2,Na V 1.3和Na ...

Efficient Transient Gene Knock-down in Tobacco Plants Using Carbon Nanocarriers
Author:
Date:
2021-01-05
[Abstract]  Gene knock-down in plants is a useful approach to study genotype-phenotype relationships, render disease resistance to crops, and enable efficient biosynthesis of molecules in plants. Small interfering RNA (siRNA)-mediated gene silencing is one of the most common ways to achieve gene knock-down in plants. Traditionally, siRNA is delivered into intact plant cells by coding the siRNA sequences into DNA vectors, which are then delivered through viral and/or bacterial methods. In this protocol, we provide an alternative direct delivery method of siRNA molecules into intact plant cells for efficient transient gene knock-down in model tobacco plant, Nicotiana benthamiana, leaves. Our approach uses one dimensional carbon-based nanomaterials, single-walled carbon nanotubes (SWNTs), to ... [摘要]  [摘要]植物基因敲低是研究基因型与表型关系,提高作物对病害的抵抗力以及实现植物分子高效生物合成的有用方法。小干扰RNA(siRNA)介导的基因沉默是在植物中实现基因敲低的最常见方法之一。传统上,通过将siRNA序列编码到DNA载体中,将siRNA传递到完整的植物细胞中,然后通过病毒和/或细菌方法传递。在这个协议中,我们提供的siRNA分子的替代直接递送方法为完整的植物细胞的高效瞬时根Ë击倒在模型的烟草植物,烟草本塞姆氏烟草,叶子。我们的方法使用一维碳基纳米材料,单壁碳纳米管(SWNTs)来传递siRNA,而不依赖于病毒/细菌的传递。我们方法的独特优势在于:i )不需要对siRNA序列进行DNA编码; ii)与非生物方法相比,这种非生物方法可在更广泛的植物物种中起作用,并且iii)使用非生物递送时,调节并发症更少方法,其中基因沉默是瞬时的,而无需对植物基因组进行永久性修饰。

图形摘要:

图形抽象标题

[背景技术[ 0002 ]在1990年代初,植物研究人员研究矮牵牛花的着色发现了通过RNA干扰(RNAi)引起的基因沉默(Van der ...

Comments