• 产品编号:FLEPO 100-2
    • 产品名称:Fluoro EPO™Fluorescent Eosinophil Peroxidase Detection Kit/荧光嗜酸性粒细胞过氧化物酶检测试剂盒
    • 产品规格:100Test
    • 价  格:6375.00
    • 供 应 商:Cell Tech
    • 库  存:
  • Fluoro EPO™
    Fluorescent Eosinophil Peroxidase 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
    Eosinophil peroxidase (EPO) is the most abundant enzyme found in eosinophils. It is the major cytotoxic agent released by activated eosinophils and uses hydrogen peroxide to generate reactive oxidants from halides and pseudo halide thiocyanate [1,2]. Eosinophils peroxidase has been shown to have antimycobacterial activity [3], however it is also implicated in tissue damage that occurs in asthma and other diseases [4,5]. Currently, the function of eosinophil involvement in the immune response is being redefined. Once considered a cell involved in host protection of parasitic infection, eosinophils multiple functions as leukocytes involved in the initiation and propagation of diverse inflammatory responses are being investigated. Eosinophils are further involved as modulators of innate and adaptive immunity [6].
    H2O2 + Detection reagent (non-fluorescent)+ EPO
                                                 ---------> fluorescent analog 
                                         Excitation:530-571nm; Emission: 590-600nm
    Applications
    • Detection of EPO activity in isolated eosinophils.
    • Functional studies of eosinophil degranulation.
    • In-vitro eosinophil chemotaxis studies.
    References
    1. Mayeno, A. N., Curran, A. J., Roberts, R. L. and Foote, C. S. (1989) Eosinophils preferentially use bromide to generate halogenating agents. J. Biol. Chem. 264, 5660±5668 .
    2. Slungaard, A. and Mahoney, J. R. (1991) Thiocyanate is the major substrate for eosinophil peroxidase in physiologic ¯uids. J. Biol. Chem. 266, 4903±4910 (Pruitt, K. M. and Tenovuo, J. V., eds.), pp. 31±53, Marcel Dekker, New York
    3. Human Eosinophil Peroxidase Induces Surface Alteration, killing, and lysis of Mycobacterium tuberculosis", Infection and Immunity, Feb. 2003, p. 605-613 by The American Society for Microbiology.
    4. Bousquet, J., Chanez, P., Lacoste, J. Y., Barneon, G., Ghavanian, N., Enander, I., Venge, P., Ahlstedt, S., Simony-Lafontaine, J., Godard, P. and Michel, F. (1990) Eosinophilic in¯ammation in asthma. New Engl. J. Med. 323, 1033±1039
    5. Parrillo, J. E., Borer, J. S., Henry, W. L., Wolff, S. M. and Fauci, A. S. (1979) The cardiovascular manifestations of the hypereosinophilic syndrome. Prospective study of 26 patients, with review of the literature. Am. J. Med. 67, 572-582
    6. Marc E. Rothenberg and ­Simon P. Hogan.­. THE EOSINOPHIL Annual Review of Immunology, Vol. 24: 147-174 (Volume publication date April 2006.)
    7. 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).
    8. 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).
    9. 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).
    10. 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).  
    11. Samantha C. Richer and W.C.L. Ford. A critical investigation of NADPH oxidase activity in human spermatozoa. Mol Hum Reprod 7, 237 (2001).
    12. William G. Gutheil, Miglena E. Stefanova and Robert A. Nicholas. Fluorescent Coupled Enzyme Assays for Image-Alanine: Application to Penicillin-Binding Protein and Vancomycin Activity Assays.  Anal Biochem 287, 196 (2000).
    13. 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).
    14. 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).
    15. D. W. R. Hall, Bridget W. Logan and G. H. Parsons. Further studies on the inhibition of monoamine oxidase by M & B 9302 (clorgyline)—I .Substrate specificity in various mammalian species. Biochemical Pharma
     
    Kit contents (for 100 assays)
    1. 5X Reaction Buffer: 60ml   ..........................................Part 3011
    2. Detection reagent: One vial for 100 assays....................Part 4007
    3. Hydrogen Peroxide: 1000µL of a stabilized 3% solution...Part 3012
    4. Eosinophil Peroxidase: 100µL in 1 vial at 10 units/ml......Part 6016
    The following kits are available:
    Catalog #
    Size
    Price (RMB)
    FLEPO 100-2
     
    100 Tests 6375.00

    Fluoro EPOFluorescent Eosinophil Peroxidase Detection Kit/荧光嗜酸性粒细胞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:

    嗜酸性粒细胞过氧化物酶(EPO)是嗜酸性粒细胞中含量最丰富的酶。它是一种由活化的嗜酸性粒细胞所释放的主要细胞毒性试剂,利用过氧化氢,从卤化物和硫氰酸盐卤化物[1,2]中生成活性氧化剂。嗜酸性粒细胞过氧化物酶已被证明具有抗分枝杆菌的活性[3],但它也存在于哮喘和其他疾病[4,5]中的组织损伤中。目前,嗜酸性粒细胞参与免疫反应的功能正在被重新定义。曾经认为有一种细胞参与保护宿主免受寄生虫的感染,嗜酸性粒细胞的多种功能中,比如白细胞参与启动和传播各种炎症反应,正在被验证。嗜酸性粒细胞作为一种调节剂可以进一步参与先天性和适应性免疫反应 [6]

     

    H2O2 + 检测试剂 (非荧光)+ EPO---------> 荧光产物

    激发波长:530-571nm; 发射波长: 590-600nm

    应用:

    在嗜酸性粒细胞中检测EPO的活性。

    嗜酸性粒细胞脱颗粒的功能研究。

    体外嗜酸性粒细胞的趋化研究。