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SpectraMax M Series Multi-Mode Microplate Readers

SpectraMax M Series Multi-Mode Microplate Readers

Company: Molecular Devices
Catalog#: M2SpectraMax
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Determination of H2O2 Generation by pHPA Assay
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2016-11-20
[Abstract]  The production of reactive oxygen species, including H2O2, is a process that can be used in signaling, cell death, or immune response. To quantify oxidative stress in cells, a fluorescence technique has been modified from a previously described method to measure H2O2 release from cells (Panus et al., 1993; Murthy et al., 2010; Larson-Casey et al., 2016; Larson-Casey et al., 2014; He et al., 2011). This assay takes advantage of H2O2-mediated oxidation of horseradish peroxidase (HRP) to Complex I, which, in turn, oxidizes p-hydroxyphenylacetic acid (pHPA) to a stable, fluorescent pHPA dimer (2,2'-dihydroxy-biphenyl-5,5’ diacetate [(pHPA)2]). The H2O2-dependent ... [摘要]  包括H 2 O 2 O 2的活性氧物质的产生是可用于信号传导,细胞死亡或免疫应答的过程。为了量化细胞中的氧化应激,已经从先前描述的方法修改了荧光技术以测量来自细胞的H 2 O 2 O 2释放(Panus等人, 2011; Larson-Casey等人,1993; Murthy等人,2010; Larson-Casey等人,2016; Larson-Casey等人 2014; He 等人,2011)。该测定利用了辣根过氧化物酶(HRP)对配合物I的H 2 O 2 O 2 - 介导的氧化,这又将对羟基苯乙酸(pHPA)氧化成稳定的荧光pHPA二聚体(2,2'-二羟基 - 联苯-5,5'-二乙酸酯[(pHPA)2])。 H 2 O 2 O 2依赖性HRP介导的pHPA氧化是用于定量H 2 O 2 O 2的灵敏和特异性的测定,/sub>从细胞释放。该测定可以测量来自全细胞,线粒体或NADPH氧化酶的H 2 O 2亚型释放。

[背景] (O 2→O 2 - )的超导阴离子(O 2 2-)的歧化导致的强烈的H 2 O 2 O 2 - 非 - 酶促反应,其中所述反应混合物是非酶促的。与O 2 - 不同,H 2 O 2可以容易地穿过膜,因此其能够氧化多个分子。 ...

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