• 产品编号:FLMPO 100-3
    • 产品名称:Fluoro MPO™ Fluorescent Myeloperoxidase Detection Kit/髓过氧化物酶检测试剂盒
    • 产品规格:500Test
    • 价  格:6375.00
    • 供 应 商:Cell Tech
    • 库  存:
  • Fluoro MPO™
    Fluorescent Myeloperoxidase Detection Kit
     
    Key Benefits:
    • Can monitor multiple time points to follow kinetics.
    • One-step, no wash assay.
    • Adaptable for High Throughput format
    • Highly Sensitive
    • Applications - Fluorescence (Plate Reader) or Absorbance
    Assay Principle
    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 powerfull bactericidal produced by neutrophils (4 review). Excessive production of these radicals can cause oxidative stress leading to oxidative tissue injury.
    H2O2 + Detection reagent (non-fluorescent)+ MPO
                                                 ---------> fluorescent analog 
                                         Excitation:530-571nm; Emission: 590-600nm
     
    Applications
    • Detection of MPO activity in neutrophils and macrophages.
    • Detection of PMN infiltration in tissue samples (inflammation and innate host defense mechanisms).  
    • Acute and chronic inflammatory disorders due to oxidative tissue damage.  
    • MPO activity in acute and chronic manifestations of cardiovascular disease.
    Citations
    Use of a hanging-weight system for liver ischemic preconditioning in mice - Hart, Much, Kohler et.al - Am J Physiol Gastrointest Liver Physiol 294: G1431-G1440, 2008.
    References
    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. Mingjie Zhou, Zhenjun Diwu, Nataliya Panchuk-Voloshina and Richard P. Haugland. A Stable Nonfluorescent Derivative of Resorufin for the Fluorometric Determination of Trace Hydrogen Peroxide: Applications in Detecting the Activity of Phagocyte NADPH Oxidase and Other Oxidases.  Anal Biochem 253, 162 (1997).
    7. J. G. Mohanty, Jonathan S. Jaffe, Edward S. Schulman and Donald G. Raible. A highly sensitive fluorescent micro-assay of H2O2 release from activated human leukocytes using a dihydroxyphenoxazine derivative. J. Immunol Methods 202, 133 (1997).
    8. Tatyana V. Votyakova and Ian J. Reynolds. Membrane Potential dependent and -independent production of reactive oxygen species by rat brain mitochondria. J Neurochem 79, 266 (2001).
    9. Chun Song, Abu B. Al-Mehdi, and Aron B. Fisher.  An immediate endothelial cell signaling response to lung ischemia.  Am J Physiol Lung Cell Mol Physiol 281, L993 (2001).  
    10. Samantha C. Richer and W.C.L. Ford. A critical investigation of NADPH oxidase activity in human spermatozoa. Mol Hum Reprod 7, 237 (2001).
    11. William G. Gutheil, Miglena E. Stefanova and Robert A. Nicholas. Fluorescent Coupled Enzyme Assays for -Alanine: Application to Penicillin-Binding Protein and Vancomycin Activity Assays.  Anal Biochem 287, 196 (2000).
    12. Dominik Peus, Remus A. Vasa, Astrid Beyerle, Alexander Meves, Carsten Krautmacher and Mark R. Pittelkow. UVB Activates ERK1/2 and p38 Signaling Pathways via Reactive Oxygen Species in Cultured Keratinocytes. J Invest Dermatol 112, 751 (1999).
    13. Tatyana V.Votyakova  ,Ian J.Reynolds. Detection of hydrogen peroxide with Amplex Red:interference by NADH and reduced glutathione auto-oxidation. Archives of Biochemistry and Biophysics, 431: 138-144 (2004).
    14. Elizabeth Forbes, Tosei Murase, Ming Yang, Klaus I. Matthaei,*James J. Lee, Nancy A. Lee,  Paul S. Foster, and Simon P. Hogan. Immunopathogenesis of Experimental Ulcerative Colitis Is Mediated by Eosinophil Peroxidase. The Journal of Immunology, 2004, 172:5664–5675.
    Kit contents (for 500 assays)
    1. 10X Assay Buffer: 60ml 
    2. Detection reagent: One vial for 500 assays.
    3. Hydrogen Peroxide: 1000µL of a stabilized 3% solution.
    4. Myeloperoxidase: 1 vial at 30 units/ml
    The following kits are available:
    Catalog #
    Size
    Price (RMB)
    FLMPO 100-3
     
    500 Tests 6375.00

    Fluoro MPO Fluorescent Myeloperoxidase Detection Kit/荧光髓过氧化物酶检测试剂盒

    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:荧光酶标仪或者吸光度仪。

    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中,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可以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。

     

    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。

    炎症慢性心血管疾病中MPO的活性。