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MTT reagent (Thiazolyl Blue Tetrazolium Bromide)


Company: Sigma-Aldrich
Catalog#: M2128
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A Modified Chromogenic Assay for Determination of the Ratio of Free Intracellular NAD+/NADH in Streptococcus mutans
[Abstract]  Nicotinamide adenine dinucleotide is a coenzyme present in all kingdoms of life and exists in two forms: oxidized (NAD+) and reduced (NADH). NAD(H) is involved in a multitude of essential metabolic redox reactions, providing oxidizing or reducing equivalents. The ratio of free intracellular NAD+/NADH is fundamentally important in the maintenance of cellular redox homeostasis (Ying, 2008). Various chromogenic cycling assays have been used to determine the ratio of NAD+/NADH in both bacterial and mammalian cells for more than forty years (Bernofsky and Swan, 1973; Nisselbaum and Green, 1969).

Here, we describe in detail an assay to determine the ratio of free intracellular NAD+ to NADH in Streptococcus mutans. This cycling ...
[摘要]  烟酰胺腺嘌呤二核苷酸是存在于生命的所有王国中的辅酶,并且以两种形式存在:氧化(NAD +)和还原(NADH)。 NAD(H)参与多种必需的代谢氧化还原反应,提供氧化或还原等价物。游离细胞内NAD + /NADH的比率在维持细胞氧化还原稳态中是根本重要的(Ying,2008)。已经使用各种显色循环测定来测定细菌和哺乳动物细胞中NAD +/NAD +/NADH的比率超过四十年(Bernofsky和Swan,1973; Nisselbaum和Green,1969)。 />  在这里,我们详细描述了测定变异链球菌中游离细胞内NAD + 与NADH的比率的测定法。该循环测定是Bernofsky和Swan(Bernofsky和Swan,1973)首先描述的方案的修改版本,使用Frezza等人描述的提取缓冲液。 (2011),随后是由Gibbon和Larher描述的减少的MTT降水(Gibon和Larher,1997)。如图1所示,使用乙醇脱氢酶来驱动一系列氧化还原反应,利用来自样品提取物的外源添加的乙醇和NAD +作为初始底物,吩嗪乙硫酸盐(PES)作为电子载体,以及噻唑基蓝色四唑溴化物(MTT)作为末端电子受体。使用6M ...

Manganese Cytotoxicity Assay on Hippocampal Neuronal Cell Culture
[Abstract]  Compared to an in vivo experiment, neuronal cell cultures are immediately accessible to observation and manipulation. In this protocol, we describe a technique to evaluate the cytotoxicity of a metal, manganese (Mn2+), on hippocampal neuronal cell cultures. Interestingly, this protocol is easily adaptable to any type of primary culture (e.g., cortical neurons) and any type of toxic compound (e.g., chemical product).

This protocol is similar to "Neuron-enriched Cultures (Method 2)" protocol (Gao, 2011).
[摘要]  与体内实验相比,神经元细胞培养物可立即进行观察和操作。 在该协议中,我们描述了评价金属锰(Mn 2 + )对海马神经元细胞培养物的细胞毒性的技术。 有趣的是,该方案容易适用于任何类型的原代培养物(例如,皮层神经元)和任何类型的有毒化合物(例如,化学产品)。
此协议类似于"神经元富集培养物(方法2)"方案(Gao,2011) 。