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Multiparameter flow cytometric analysis of Myeloperoxidase expression in Human peripheral blood leucocytes. Human whole blood was treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to remove erythrocytes and to fix leucocytes. The fixed leucocytes were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) and then stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human Myeloperoxidase antibody (Cat. No. 568365/568366). The bivariate pseudocolor density plot showing the correlated expression of Myeloperoxidase (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-Human Myeloperoxidase
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Reacts with myeloperoxidase (MPO), a hemoprotein of 140 kDa, composed of two heavy subunits of 52 kDa and two light chains of 15 kDa. MPO is stored in primary azurophilic granules of neutrophils and plays a major role in the bactericidal activity of neutrophils during phagocytosis. It catalyzes the generation of hypochlorous acid, a powerful oxidant. 1B10 antibody detects MPO in neutrophils, monocytes and HL-60 cells. This reagent is suitable for immunohistochemical staining of acetone-fixed, frozen tissue section, or formalin-fixed, paraffin-embedded tissue sections with TUF pretreatment, or zinc-fixed, paraffin-embedded tissue sections with no pretreatment.
Development References (3)
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Audrain MA, Baranger TA, Moguilevski N, et al. Anti-native and recombinant myeloperoxidase monoclonals and human autoantibodies. Clin Exp Immunol. 1997; 107(1):127-134. (Immunogen: ELISA, Western blot). View Reference
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Kennedy AD, Willment JA, Dorward DW, Williams DL, Brown GD, DeLeo FR. Dectin-1 promotes fungicidal activity of human neutrophils.. Eur J Immunol. 2007; 37(2):467-78. (Clone-specific: Western blot). View Reference
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Lanza F, Latorraca A, Moretti S, Castagnari B, Ferrari L, Castoldi G. Comparative analysis of different permeabilization methods for the flow cytometry measurement of cytoplasmic myeloperoxidase and lysozyme in normal and leukemic cells. Cytometry. 1997; 30(3):134-144. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.