中文名称

Class III HDAC, BioAssay™

英文名字

Class III HDAC, BioAssay™

供应商

USBiological

产品货号

029446

产品报价

￥1/96Tests

产品说明书

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购买方式

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产品新闻

背景资料

USBiological品牌是全球有名的抗原抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。武汉艾美捷科技作为USBiological品牌中国区域总代理，是行业中少有的致力于服务客户，帮助客户，且拥有独立的专业销售团队、技术支持团队、市场营销团队、进出口报关团队的高科技生物企业。可以为您提供及时的咨询响应，专业的产品和解决方案支持，稳健快捷的交货周期，优质放心的售后服务。我们致力于为您提供有价值的产品和服务，在意您的成功！

产品描述

Posttranslational acetylation of lysines occurs in a wide variety of proteins, and consequently regulates a diverse array of intermolecular interactions and biological processes. Proteomic analysis of lysine acetylation in mammalian cells has revealed >3600 acetylated lysine sites occurring on >1700 proteins. This diverse array of proteins function in many major cellular processes, including gene regulation, cytoskeletal organization, protein trafficking, and cell signaling. On histone proteins, the universe of lysine acetylation is controlled by the opposing enzymatic activities of histone acetyltransferases (HAT) and histone deacetylases (HDAC). Given the diversity of lysine acetylated proteins and processes in which they occur, modulators of HATs and HDACs are major targets for drug discovery efforts in a number of disease processes, including metabolism, cancer and cardiovascular disease.Four different classes of histone deacetylases have been described. The first two classes are HDACs which are inhibited by trichostatin A (TSA). The third group (Class III HDAC's) is NAD+-dependent proteins not inhibited by TSA. The fourth classes of HDACs are atypical and are considered HDAC's due to DNA sequence similarity to the other classes of HDAC's.Class III HDAC's, also known as sirtuins, are mechanistically distinct from class I and class II HDACs in that they couple deacetylation of the peptide/protein substrate to cleavage of NAD+ to form nicotinamide and O-acetyl-ADP-ribose. The first member of the sirtuin family to be assigned a physiological role was the yeast Sir2, which was shown to increase mother cell life span when over expressed. The mammalian sirtuins comprise 7 members, termed Sirt1-7, which share the NAD+-binding catalytic domain, but differ in N- and C-termini, subcellular localization, substrate preference, and ultimately in biological function. For example, Sirt1 resides primarily in the nucleus, where it deacetylates several transcriptional regulators, including PGC-1a to promote mitochondrial biogenesis and gluconeogenesis, and p53 to reduce apoptosis. Sirtuins 3-5 are located in mitochondria, and Sirt3 has been shown to deacetylate several metabolic enzymes to divert alternative carbon sources to the TCA cycle.Multiple lines of evidence have suggested that the activation of sirtuins in vivo could represent therapeutic target for the maintenance of metabolic homeostasis. Subsequent studies searching for activators of sirtuin activity indicated the plant polyphenol resveratrol as a potential activator of Sirt1. However, these studies were called into question as a result of data demonstrating that resveratrol-mediated activation of sirtuins was actually due to an artifact of the fluorescently labeled peptide substrate used in the commercially available assay used for these studies.The SIRTainty approach eliminates artifacts due to resveratrol and potential interference of enzyme substrate interactions due to the presence of bulky fluorophores. Unlike other sirtuin assays that utilize a fluorescently tagged acetylated peptide substrate, the SIRTainty assay employs an untagged acetylated peptide substrate. This approach not only enables unparalleled flexibility in your choice of sirtuin isoform and peptide substrate, but also helps avoid potential artifacts attributed to the use of a fluorescently labeled substrate.Intended Use:For measuring sirtuin activity as well as for screening of activators and inhibitors of sirtuin enzymes.Specificity:Class III HDACTest Principle:To perform this assay, a sirtuin enzyme, beta-NAD, acetylated peptide substrate, test compound, and nicotinamidase enzyme are combined and incubated for 30 minutes. During this time the acetylated peptide substrate is acted upon by the sirtuin enzyme to produce nicotinamide. In a secondary reaction, the nicotinamidase enzyme converts the nicotinamide into nicotinic acid and NH3+ (free ammonia). To generate a signal for readout, a proprietary developer reagent is added and the signal is read using a fluorescent plate reader.Kit Components:Black plate: 1x96 wellsAssay Buffer: 1x10mlPlate Covers: 2 plate coversAcetylated Peptide Substrate: 1x75ul (1mM)beta NAD: 1x30ul (50mM)Nicotinamide (NAM): 1x10ul (10mM)Recombinant Sirt1 Enzyme: 1x120 unitsRecombinant Nicotinamidase Enzyme with 50% glycerol: 1x60ug (1mg/ml)Suramin (Sirt Inhibitor) in ddH2O: 1x150ul (25mM)Developer Reagent: 1x4ml (10mM)Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

产品形式

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保存建议

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其他

Usbiological公司是美国著名的抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。Usbiological公司现已拥有超过50,19234种抗体、抗原和生化产品，为科研用户提供了诸多超值选择。

注意

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