Description: The monoclonal antibody MPO455-8E6 recognizes myeloperoxidase (MPO), a protein within the azurophilic granules of myeloid cells. MPO is a multimeric protein comprised of two 55 kDa subunits and two 15 kDa subunits. The larger subunits associate with a heme protein resulting in a greenish color. As an enzyme, MPO breaks down hydrogen peroxide and oxidizes tyrosine. The products of this reaction, hypochlorous acid and tyrosyl radical, cause the cytotoxic and killing effects characteristic of neutrophils. Myeloperoxidase is important in the diagnosis of some cancers and increases in serum levels have been shown to correlate with cardiac events. Applications Reported: This MPO455-8E6 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This MPO455-8E6 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells using the Intracellular Fixation & Permeablization Buffer Set (cat. 88-8824). For best results, whole blood should first be stained with antibodies to surface antigens then treated with 1X RBC lysis buffer (cat. 00-4333) to lyse erythrocytes. Then, for intracellular staining for MPO, cells should be fixed with IC Fixation Buffer (cat. 00-8222) washed two times with Permeabilization Buffer (cat. 00-8333) and then incubated with MPO455-8E6 for 30-60 minutes.After washing, cells may be analyzed on a flow cytometer. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered. Myeloperoxidase (MPO) is a hemoprotein that is abundantly expressed in neutrophils and secreted during their activation. Native Myeloperoxidase is represented as a covalently bound tetrameric complex of two glycosylated alpha chains with a molecular weight range of 59 - 64 kDa, and two unglycosylated beta chains (MW ~14 kDa) with total MW ~150 kDa and a theoretical pI of 9.2. Traditionally, myeloperoxidase was considered as a main target of anti-neutrophil cytoplasm antibodies (ANCA), the serological markers for certain systemic vasculities such as periarteriitis nodosa, microscopic polyarteriitis and pulmonary eosinophilic granulomatosis (Churg-Strauss syndrome). Low to moderate anti-myeloperoxidase autoantibody levels are also reported in rheumatoid arthritis. Recently, it was shown that myeloperoxidase participates in the initiation and progression of cardiovascular disease. Myeloperoxidase possesses potent proinflammatory properties and may contribute directly to tissue injury. Now Myeloperoxidase is under consideration as one of the most promising cardiac markers. Myeloperoxidase is part of the host defense system of polymorphonuclear leukocytes and myeloperoxidase is responsible for microbicidal activity against a wide range of organisms. In the stimulated PMN (polymorphonuclear leukocytes), MPO catalyzes the production of hypohalous acids, primarily hypochlorous acid in physiologic situations, and other toxic intermediates that greatly enhance PMN microbicidal activity.
|4° C, store in dark, DO NOT FREEZE!|
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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