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Fluoro MPOHOCL™
Fluorescent Myeloperoxidase Chlorination and peroxidation Detection Kit New! |
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| Assay Principle | |||||||||
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Myeloperoxidase (MPO) is a highly cationic glycosolated hemoprotein that has a molecular weight of 144kD. The hemoprotein consists of two dimers linked via a disulfide bridge. Each dimmer is composed of a heavy (53kD) and light (15kD) subunit. Each heavy chain contains an independently acting protoporphyrin group containing a central iron (1-5). MPO is present in the azurophilic granules of polymorphonuclear leukocytes (PMNs) and is unique to neutrophils and monocytes. However, monocytes contain only one third of the MPO found in PMN’s. MPO utilizes H202 produced by the neutrophils to oxidize a varity of aromatic compounds to give substrate radicals for bactericidal activity (4 review). This enzyme is unique however in that it can oxidize chloride ions to produce a strong nonradical oxidant,HOCl. HOCl is the most powerful bactericidal produced by neutrophils (4 review). Excessive production of these radicals can cause oxidative stress leading to oxidative tissue injury.
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| Chlorination Reaction:
H2O2 + MPO + Cl- ----> HOCl + APF (non fluorescent) ----> Fluorescent Dye
Excitation:488nm; Emission: 515-530nm Peroxidation Reaction:
H2O2 + Detection reagent (non-fluorescent)+ MPO -----> fluorescent analog
Excitation 530-571nm Emission 590-600nm
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| References | |||||||||
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1.Olsen, R. L. & Little, c. (1983) Biochem. J. 209, 781-787.
2.Nauseef, W. M., and Malech, H. L. (1986) Blood, 67, 1504-1507.
3. Andrews, P. C., Parnes, C., and Krinsky, N. I. (1984) Arch.
Biochem. Biophys., 228, 439-442.
4. Mark B. Hampton, Anthony J. Kettle, and Christine C. Winterbourn. Inside the Neutrophil Phagosome: Oxidants, Myeloperoxidase, and Bacterial Killing. Blood, Vol. 92 No. 9 (November 1), 1998: pp. 3007-3017.
5. Andrews, P.C., Parnes, C. & Krinsky, N.I. (1984) Comparison of myeloperoxidase and hemi-myeloperoxidase with respect to catalysis, regulation, and bacterial activity. Arch. Biochem. Biophys. 228, 439–442.
6. Ken-ichi Setsukinai, Yasuteru Urano, Katsuko Kakinuma, Hideyuki J. Majima , and Tetsuo Nagano. Development of Novel Fluorescence Probes That Can Reliably Detect Reactive Oxygen Species and Distinguish Specific Species. THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 278, No. 5, Issue of January 31, pp. 3170–3175, 2003
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| Kit contents (for 500 assays of Myeloperoxidase detection and 300 assays of Chlorination detection) | |||||||||
Not For Sale in Japan
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Fluoro MPOHOCL™ Fluorescent Myeloperoxidase Chlorination and peroxidation Detection Kit/荧光髓过氧化物酶氯化法和过氧化反应检测试剂盒
主要优点：
遵循动力学，可以检测多个时间点。
一步法，免洗。
适用于高通量模式。
高灵敏性。
应用：荧光酶标仪。
检测原理：
髓过氧化物酶（MPO）是一种高度阳离子糖基化红血素蛋白，分子量是144kD。该红血素蛋白由两个通过二硫键连接而成的两个二聚体组成。每个亚单位是由一个重亚基（53kD）和一个轻亚基（15kD）组成。每个重链包含一个独立的原卟啉组分（包含一个中央铁离子）（1-5）。 MPO存在于中性粒细胞中的嗜苯胺蓝颗粒，并且对中性粒细胞和单核细胞有唯一性。然而，单核细胞中只含有中性粒细胞的过氧化物酶含量的三分之一。 MPO利用嗜中性粒细胞产生的H2O2氧化各种芳香族化合物，给杀菌活性提供基础（报告4）。然而，这种酶是独一无二的，它可以氧化氯离子，产生强烈的嗜中性粒细胞氧化产物，次氯酸。这些自由基的过量产生会导致氧化应激，导致组织损伤。
氯化反应:
H2O2 + MPO + Cl- ----> HOCl + APF (非荧光) ----> 荧光染料
激发波长:488nm; 发射波长: 515-530nm
过氧化反应:
H2O2 +检测试剂 (非荧光)+ MPO -----> 荧光类似物
激发波长： 530-571nm 发射波长： 590-600nm
应用：
检测嗜中性粒细胞和巨噬细胞中的MPO活性。
检测组织样本中多形核白细胞的渗透（炎症和宿主的防御机制）
由于氧化造成组织损伤，带来的严重、慢性炎症紊乱。
检测炎症慢性心血管疾病中MPO的活性。
