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      RB780 Mouse Anti-Human IL-1ra
      RB780 Mouse Anti-Human IL-1ra
      Multiparameter flow cytometric analysis of IL-1ra expression in stimulated Human PBMC.  Human PBMC were cultured with Recombinant Human IFN-γ protein (Cat. No. 554617 at 20 ng IFN-γ/ml for 2h; 37°C) and then lipopolysaccharide (LPS) [1 µg LPS/ml for 24 h; 37°C] was added with Protein Transport Inhibitor (BD GolgiStop™) [Cat. No. 554724]. Cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained in BD Perm/Wash™ Buffer with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human IL-1ra antibody (Cat. No. 571512/571562; Right Plot). The pseudocolor density plot showing the correlated expression of IL-1ra (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 PBMC. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
      RB780 Mouse Anti-Human IL-1ra
      Multiparameter flow cytometric analysis of IL-1ra expression in stimulated Human PBMC.  Human PBMC were cultured with Recombinant Human IFN-γ protein (Cat. No. 554617 at 20 ng IFN-γ/ml for 2h; 37°C) and then lipopolysaccharide (LPS) [1 µg LPS/ml for 24 h; 37°C] was added with Protein Transport Inhibitor (BD GolgiStop™) [Cat. No. 554724]. Cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained in BD Perm/Wash™ Buffer with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human IL-1ra antibody (Cat. No. 571512/571562; Right Plot). The pseudocolor density plot showing the correlated expression of IL-1ra (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 PBMC. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
      Product Details
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      BD Horizon™
      IL-1RN; IL-1ra; IL1 inhibitor; IL1F3; IL1RN; IRAP; sIL-1ra; icIL-1ra
      Human (QC Testing)
      Mouse BALB/c IgG1, κ
      Recombinant Human IL-1 Receptor Antagonist Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl/test
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. For U.S. patents that may apply, see bd.com/patents.
      5. 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.
      6. 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).
      7. 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.
      8. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      11. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      12. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
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      571562 Rev. 1
      Antibody Details
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      AS17

      The AS17 monoclonal antibody specifically recognizes the Interleukin-1 receptor antagonist (IL-1ra or IL-1Ra) which is also known as IL-1 receptor antagonist protein (IRAP), IL-1F3, IL-1RN, and IL1 inhibitor. IL-1ra is an ~17-25 kDa glycoprotein that is encoded by IL1RN. Variants of IL-1ra include a secreted form, sIL-1ra, that is produced by a variety of cell types including activated monocytes, macrophages, neutrophils, and keratinocytes. Intracellular variants of IL-1ra (icIL-1ra) that remain in the cytoplasm have also been described. These include icIL-1Ra1 that is expressed in keratinocytes and other epithelial cells, endothelial cells, fibroblasts, and macrophages. The icIL-1Ra3 variant is expressed in neutrophils and hepatocytes. icIL-1ra proteins may become functional after their release from dead or dying cells although there are indications for their functions inside cells as well. IL-1ra binds to Interleukin-1 Receptor type I (IL-1R1, also known as CD121a) and prevents IL-1α or IL-1β from binding to IL-1R1. IL-1R1 is part of the heterodimeric IL-1 Receptor signaling complex that is comprised of IL-1R1 and the IL-1R accessory protein (IL-1RAcP). icIL-1Ra1 binds to IL-1R1 avidly whereas icIL-1Ra3 binds weakly. IL-1ra does not bind to IL-1RAcP. IL-1ra also binds with low affinity to the nonsignaling Interleukin-1 Receptor type II (IL-1R2, also known as CD121b). IL-1R2 also helps regulate IL-1-mediated activities by acting as a decoy receptor for IL-1 on the membrane of cells or in a soluble form. IL-1ra plays key regulatory roles in health as well as in a variety of diseases including sepsis, colitis, and rheumatoid arthritis.

      571562 Rev. 1
      Format Details
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      RB780
      The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
      altImg
      RB780
      Blue 488 nm
      498 nm
      781 nm
      571562 Rev.1
      Citations & References
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      View product citations for antibody "571562" on CiteAb

      Development References (12)

      1. Arend WP, Joslin FG, Thompson RC, Hannum CH. An IL-1 inhibitor from human monocytes. Production and characterization of biologic properties.. J Immunol. 1989; 143(6):1851-8. (Biology). View Reference
      2. Arend WP, Malyak M, Guthridge CJ, Gabay C. Interleukin-1 receptor antagonist: role in biology.. Annu Rev Immunol. 1998; 16:27-55. (Biology). View Reference
      3. Arend WP, Palmer G, Gabay C. IL-1, IL-18, and IL-33 families of cytokines.. Immunol Rev. 2008; 223:20-38. (Clone-specific). View Reference
      4. Dinarello CA, van der Meer JW. Treating inflammation by blocking interleukin-1 in humans.. Semin Immunol. 2013; 25(6):469-84. (Biology). View Reference
      5. Dinarello CA. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases.. Blood. 2011; 117(14):3720-32. (Biology). View Reference
      6. Hannum CH, Wilcox CJ, Arend WP, et al. Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor.. Nature. 1990; 343(6256):336-40. (Biology). View Reference
      7. Haskill S, Martin G, Van Le L, et al. cDNA cloning of an intracellular form of the human interleukin 1 receptor antagonist associated with epithelium.. Proc Natl Acad Sci U S A. 1991; 88(9):3681-5. (Biology). View Reference
      8. Janson RW, Hance KR, Arend WP. Production of IL-1 receptor antagonist by human in vitro-derived macrophages. Effects of lipopolysaccharide and granulocyte-macrophage colony-stimulating factor.. J Immunol. 1991; 147(12):4218-23. (Biology). View Reference
      9. Kennedy MC, Rosenbaum JT, Brown J, et al. Novel production of interleukin-1 receptor antagonist peptides in normal human cornea.. J Clin Invest. 1995; 95(1):82-8. (Biology). View Reference
      10. Krzesicki RF, Hatfield CA, Bienkowski MJ, et al. Regulation of expression of IL-1 receptor antagonist protein in human synovial and dermal fibroblasts.. J Immunol. 1993; 150(9):4008-18. (Biology). View Reference
      11. McColl SR, Paquin R, Ménard C, Beaulieu AD. Human neutrophils produce high levels of the interleukin 1 receptor antagonist in response to granulocyte/macrophage colony-stimulating factor and tumor necrosis factor alpha.. J Exp Med. 1992; 176(2):593-8. (Biology). View Reference
      12. Rhind SG, Castellani JW, Brenner IK, et al. Intracellular monocyte and serum cytokine expression is modulated by exhausting exercise and cold exposure.. Am J Physiol Regul Integr Comp Physiol. 2001; 281(1):R66-75. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
      View All (12) View Less
      571562 Rev. 1

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      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.