| Determination of H2O2 Generation by pHPA Assay
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Author:
Date:
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) ...
[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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| Measurement of Transferrin- and Non-transferrin-bound Iron Uptake by Mouse Tissues
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Author:
Date:
2016-09-05
[Abstract] Iron in blood plasma is bound to its transport protein transferrin, which delivers iron to most tissues. In iron overload and certain pathological conditions, the carrying capacity of transferrin can become exceeded, giving rise to non-transferrin-bound iron, which is taken up preferentially by the liver, kidney, pancreas, and heart. The measurement of tissue transferrin- and non-transferrin-bound iron (TBI and NTBI, respectively) uptake in vivo can be achieved via intravenous administration of 59Fe-labeled TBI or NTBI followed by gamma counting of various organs. Here we ...
[摘要] 血浆中的铁结合其转运蛋白转铁蛋白，其将铁递送至大多数组织。 在铁过载和某些病理状况下，转铁蛋白的携带能力可能超过，导致非转铁蛋白结合的铁，其优先被肝脏，肾脏，胰腺和心脏摄取。 分别测量组织转铁蛋白和非转铁蛋白结合的铁（分别为TBI和NTBI）在体内的摄取可以通过静脉内施用59 Fe标记的TBI或 NTBI，然后是各种器官的γ计数。 在这里我们描述了测量小鼠组织的TBI和NTBI摄取的详细协议。
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| Human Blood Component Vaccinia Virus Neutralization Assay
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Author:
Date:
2015-12-05
[Abstract] Many therapeutic viruses, such as oncolytic viruses, vaccines, or gene therapy vectors, may be administered by the intravenous route to maximize their delivery to target tissues. Blood components, such as antibody, complement and blood cells (such as neutrophils, monocytes, T cells, B cells or platelets) may result in viral neutralization and therefore reduce the therapeutic efficacy. This protocol will describe an in vitro assay by which to test the interaction of viruses with blood components. The effect of various factors can be isolated through fractionation. While whole blood ...
[摘要] 许多治疗性病毒，例如溶瘤病毒，疫苗或基因治疗载体，可以通过静脉内途径施用以最大化它们向靶组织的递送。 血液组分，例如抗体，补体和血细胞（例如嗜中性粒细胞，单核细胞，T细胞，B细胞或血小板）可导致病毒中和并因此降低治疗功效。 该方案将描述用于测试病毒与血液组分的相互作用的体外测定。 各种因素的影响可以通过分馏分离。 尽管全血可以提供最具生理学相关性的快照，但血浆可以研究抗体与补体一起的作用，而热灭活的血浆将会询问单独的抗体的作用。
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