NWLSS™ Superoxide Oxidant Status Assay推荐用于测定分离的单核细胞群中的超氧化物和其他生物样品中的一般氧化水平。
Superoxide and other reactive oxygen species play very important role in diseases and pathology related to oxidative stress. Their extremely low steady-state concentration in-vivo require sensitive methods of measurement. A number of techniques have been described in the literature including ferricytochrome c reduction, nitroblue tetrazolium reduction, aconitase activity inhibition, nitrone spin trapping, electrochemical detection, and chemiluminescent assays with luminol and lucigenin. The luminol based assay has been criticized as a specific quantitative method to determine superoxide production in cells due to its sensitivity to a number of reactive oxygen species (hypochlorite, peroxynitrite and hydroxyl radical from H2O2+metal/heme-proteins). Also, the first intermediate in the reaction can react with oxygen to generate superoxide resulting in higher apparent superoxide production rate than actual cellular production rate. However, this assay can provide a valid, simple and sensitive gross assessment of reactive oxygen species in many biological sample types and is an important method in the analysis of specific superoxide production by monocytes.
Test Principle
The NWLSS™ NWK-SOS01 method is based on Luminol reaction with superoxide to produce a luminophore with an emission peak at ~425 nm. Orthovanadate is used to increase luminescence respose by uo to 50X. The luminescence intensity is proportional to the amount of superoxide in the sample.
方法:化学发光
应用:化学发光方法,用于测定单核细胞中超氧化物水平并提供其他生物样品中一般氧化剂水平数据
样品要求:分离的细胞群、全血、组织匀浆
特异性:测量分离的单核细胞群时的超氧化物,以及生物样品中的一般氧化剂水平
灵敏度:
LLD = 0.3 U/mL 在反应混合物中
5 U/mL 在样品中添加到反应混合物中
储存和稳定性:储存于 2-8℃
试剂盒内容:
Luminol Solution
HBSS Buffer Solution
Enhancer Reagent
Phorbol Myristate Acetate (PMA)
PMA Solvent (DMSO)