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      R718 Mouse Anti-Human Neutrophil Elastase
      R718 Mouse Anti-Human Neutrophil Elastase

      Flow cytometric analysis of Human Neutrophil Elastase expression in Human peripheral blood granulocytes and tumor cell lines.  Human peripheral blood granulocytes (Left Plot) prepared by density gradient centrifugation and cells from the Human Jurkat (Acute T cell leukemia, ATCC® TIB-152™) cell line (Middle Plot) and Human U-937 (Histiocytic lymphoma, ATCC® CRL-1593™) cell line (Right Plot) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms) or BD Horizon™ R718 Mouse Anti-Human Neutrophil Elastase antibody (Cat. No. 569519/569520; solid line histograms) at 1 µg/test. The fluorescence histograms showing Neutrophil Elastase expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

      R718 Mouse Anti-Human Neutrophil Elastase

      Flow cytometric analysis of Human Neutrophil Elastase expression in Human peripheral blood granulocytes and tumor cell lines.  Human peripheral blood granulocytes (Left Plot) prepared by density gradient centrifugation and cells from the Human Jurkat (Acute T cell leukemia, ATCC® TIB-152™) cell line (Middle Plot) and Human U-937 (Histiocytic lymphoma, ATCC® CRL-1593™) cell line (Right Plot) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms) or BD Horizon™ R718 Mouse Anti-Human Neutrophil Elastase antibody (Cat. No. 569519/569520; solid line histograms) at 1 µg/test. The fluorescence histograms showing Neutrophil Elastase expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

      Product Details
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      BD Horizon™
      ELANE; HNE; PMN elastase; PMN-E; neutrophil elastase
      Human (QC Testing)
      Mouse IgG1
      Neutrophil granule proteins
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      1991
      AB_3685127
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
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      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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. 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.
      5. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
      8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      9. For U.S. patents that may apply, see bd.com/patents.
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      569519 Rev. 1
      Antibody Details
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      NP57

      The NP57 monoclonal antibody specifically recognizes human Neutrophil Elastase (NE) which is also known as Human leukocyte elastase (HLE), Granulocyte-derived Elastase (GE), or PMN Elastase (PMN-E). Neutrophil Elastase is a major serine proteinase encoded by ELANE (Elastase, neutrophil expressed) that belongs to the elastase subfamily within the peptidase S1 family. Neutrophil Elastase is predominantly present in the azurophilic granules (primary granules) of neutrophils and monocytes. When released outside of these effector cells, it can act to hydrolyze a variety of extracellular matrix substrates including elastin, fibronectin, collagen, and proteoglycans. Elastase activity in normally controlled by serine proteinase inhibitors. Neutrophil Elastase can regulate the functions of NK cells, monocytes, and granulocytes, eg, by inhibiting C5a-dependent enzyme release and chemotaxis by neutrophils. It is also capable of killing bacteria such as E. coli and degrading virulence factors from bacteria such as Yersinia, Shigella, and Salmonella. Dysregulated expression or defects in this enzyme are associated with some inflammatory and degenerative diseases including rheumatoid arthritis, emphysema, chronic obstructive pulmonary disease, and cystic fibrosis as well as neutropenias.

      569519 Rev. 1
      Format Details
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      R718
      The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      R718
      Red 627-640 nm
      695 nm
      718 nm
      569519 Rev.1
      Citations & References
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      View product citations for antibody "569519" on CiteAb

      Development References (7)

      1. Belaaouaj A, Kim KS, Shapiro SD. Degradation of outer membrane protein A in Escherichia coli killing by neutrophil elastase.. Science. 2000; 289(5482):1185-8. (Biology). View Reference
      2. Cramer EM, Beesley JE, Pulford KA, Breton-Gorius J, Mason DY. Colocalization of elastase and myeloperoxidase in human blood and bone marrow neutrophils using a monoclonal antibody and immunogold.. Am J Pathol. 1989; 134(6):1275-84. (Clone-specific: Electron microscopy, Immunofluorescence, Immunohistochemistry). View Reference
      3. Davies PL, Maxwell NC, Kotecha S, Spiller OB. Monoclonal anti-neutrophil elastase antibody characterisation: ability to block function, detect free versus serpin-complexed enzyme and stain intracellular granules.. J Immunol Methods. 2008; 336(2):175-82. (Clone-specific: Flow cytometry). View Reference
      4. Korkmaz B, Attucci S, Jourdan ML, Juliano L, Gauthier F. Inhibition of neutrophil elastase by alpha1-protease inhibitor at the surface of human polymorphonuclear neutrophils.. J Immunol. 2005; 175(5):3329-38. (Biology). View Reference
      5. Pulford KA, Erber WN, Crick JA, et al. Use of monoclonal antibody against human neutrophil elastase in normal and leukaemic myeloid cells.. J Clin Pathol. 1988; 41(8):853-60. (Immunogen: Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western blot). View Reference
      6. Tralau T, Meyer-Hoffert U, Schröder JM, Wiedow O. Human leukocyte elastase and cathepsin G are specific inhibitors of C5a-dependent neutrophil enzyme release and chemotaxis.. Exp Dermatol. 2004; 13(5):316-25. (Biology). View Reference
      7. Weinrauch Y, Drujan D, Shapiro SD, Weiss J, Zychlinsky A. Neutrophil elastase targets virulence factors of enterobacteria.. Nature. 2002; 417(6884):91-4. (Biology). View Reference
      View All (7) View Less
      569519 Rev. 1

      Please refer to Support Documents for Quality Certificates

       

      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

       

      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.