{{'Search' | translate}}
 

Trichloroacetic acid

Company: Sigma-Aldrich
Catalog#: T4885
Bio-protocol()
Company-protocol()
Other protocol()

Two Different Methods of Quantification of Oxidized Nicotinamide Adenine Dinucleotide (NAD+) and Reduced Nicotinamide Adenine Dinucleotide (NADH) Intracellular Levels: Enzymatic Coupled Cycling Assay and Ultra-performance Liquid Chromatography (UPLC)-Mass Spectrometry
Author:
Date:
2018-07-20
[Abstract]  Current studies on the age-related development of metabolic dysfunction and frailty are each day in more evidence. It is known, as aging progresses, nicotinamide adenine dinucleotide (NAD+) levels decrease in an expected physiological process. Recent studies have shown that a reduction in NAD+ is a key factor for the development of age-associated metabolic decline. Increased NAD+ levels in vivo results in activation of pro-longevity and health span-related factors. Also, it improves several physiological and metabolic parameters of aging, including muscle function, exercise capacity, glucose tolerance, and cardiac function in mouse models of natural and accelerated aging.

Given the importance of monitoring cellular NAD+ and ...
[摘要]  目前关于代谢功能障碍和虚弱的年龄相关发展的研究每天都有更多的证据。众所周知,随着衰老的进展,烟酰胺腺嘌呤二核苷酸(NAD + )水平在预期的生理过程中降低。最近的研究表明,NAD + 的减少是与年龄相关的代谢衰退发展的关键因素。增加NAD + 水平体内导致激活寿命和健康跨度相关因素。此外,它改善了老化的几个生理和代谢参数,包括自然和加速老化的小鼠模型中的肌肉功能,运动能力,葡萄糖耐量和心脏功能。

鉴于监测细胞NAD + 和NADH水平的重要性,有一个值得信赖的方法是至关重要的。该方案的目的是在有效且广泛适用的测定中描述来自组织和细胞的NAD + 和NADH提取以及其图形和定量分析。

【背景】氧化烟酰胺腺嘌呤二核苷酸和还原型烟酰胺腺嘌呤二核苷酸(NAD + 和NADH)是重要的生物辅助因子,它们在几种合成代谢和分解代谢功能中提供和接受电子。它们参与诸如糖酵解,三羧酸循环和氧化磷酸化的反应。此外,它还作为DNA损伤修复中涉及的几种酶的底物,例如sirtuins和poly(ADP-核糖)聚合酶(PARP)(Imai和Guarente,2014; Verdin,2015; Yoshino et al。 ,2018)。

NAD + ...

In vitro Assessment of RNA Polymerase I Activity
Author:
Date:
2017-02-05
[Abstract]  In eukaryotic cells transcriptional processes are carried out by three different RNA polymerases: RNA polymerase I which specifically transcribes ribosomal RNA (rRNA), RNA polymerase II which transcribes protein-coding genes to yield messenger RNAs (mRNAs) and small RNAs, while RNA polymerase III transcribes the genes for transfer RNAs and for the smallest species of ribosomal RNA (5S rRNA). This protocol describes an in vitro assay to evaluate the rRNA transcriptional activity of RNA polymerase I. The method measures the quantity of radiolabelled uridine 5’ triphosphate incorporated in ex novo synthesized rRNA molecules by RNA polymerase I, in optimal conditions for the enzyme activity and in the presence of a toxin, α-amanitin, which inhibits RNA polymerase II and III ... [摘要]  在真核细胞中,转录过程由三种不同的RNA聚合酶:特异性转录核糖体RNA(rRNA)的RNA聚合酶I,转录蛋白质编码基因以产生信使RNA(mRNA)和小RNA的RNA聚合酶II进行转录,RNA聚合酶III转录转录RNA和最小核糖体RNA(5S rRNA)的基因。该方案描述了用于评价RNA聚合酶I的rRNA转录活性的体外实验方法。该方法测量了合并的rRNA分子中掺入的放射性标记的尿苷5'-三磷酸的量通过RNA聚合酶I,在酶活性的最佳条件和毒素α-amanitin存在下,其抑制RNA聚合酶II和III而不影响RNA聚合酶I(Novello和Stirpe,1970)。

背景 在真核细胞中,RNA聚合酶I转录位于核仁中的核糖体基因,产生45S rRNA前体分子。这些被处理以形成成熟的18S,5.8S和28S ...

Comments