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      PE Rat Anti-Mouse Ly-6C
      PE Rat Anti-Mouse Ly-6C
      Multicolor flow cytometric analysis of Ly-6C expression on Mouse splenic leucocytes.     Top Plots: C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were then stained with APC Rat Anti-Mouse CD8a antibody (Cat. No. 553035) and with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse Ly-6C antibody (Cat. No. 568954; Right Plot) at 0.25 μg/test.     Bottom Plots: C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with BD Horizon™ BUV395 Rat Anti-Mouse CD11b antibody (Cat. No. 563553) and with either PE Rat IgG1, κ Isotype Control (Left Plot) or PE Rat Anti-Mouse Ly-6C antibody (Right Plot) at 0.25 μg/test.     DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of Ly-6C (or Ig Isotype control staining) versus CD8a (Top Plots) or CD11b (Bottom Plots) were derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      PE Rat Anti-Mouse Ly-6C
      Multicolor flow cytometric analysis of Ly-6C expression on Mouse splenic leucocytes.     Top Plots: C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were then stained with APC Rat Anti-Mouse CD8a antibody (Cat. No. 553035) and with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse Ly-6C antibody (Cat. No. 568954; Right Plot) at 0.25 μg/test.     Bottom Plots: C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with BD Horizon™ BUV395 Rat Anti-Mouse CD11b antibody (Cat. No. 563553) and with either PE Rat IgG1, κ Isotype Control (Left Plot) or PE Rat Anti-Mouse Ly-6C antibody (Right Plot) at 0.25 μg/test.     DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of Ly-6C (or Ig Isotype control staining) versus CD8a (Top Plots) or CD11b (Bottom Plots) were derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      Ly-6C; Ly-6C1; Ly6c; Ly6c1; Lymphocyte antigen Ly-6C
      Mouse (QC Testing)
      Rat IgG1, κ
      L3 Clone CTL Cells
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      17067
      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 unconjugated antibody and free dye were 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 cell 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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      568954 Rev. 1
      Antibody Details
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      HK1.4.rMAb

      The HK1.4.rMAb monoclonal antibody is a recombinant monoclonal antibody derived from HK1.4 hybridoma cells that specifically recognizes a non-polymorphic determinant on Mouse Lymphocyte antigen Ly-6C which is also known as Ly6c. Ly-6C is an ~14-17 kDa glycosylphosphatidylinositol (GPI)-linked cell-surface antigen that is encoded by Ly6c1 (Lymphocyte antigen 6 complex, locus C1) which belongs to the Ly-6 gene family. Ly-6C is expressed on monocytes, macrophages, neutrophils, eosinophils, endothelial cells, plasma cells, thymocytes, NK cells, and some T cell subsets. Mice with the Ly-6.2 haplotype (eg, AKR, C57BL, C57BR, C57L, C58, DBA/2, PL, SJL, SWR, 129) have subsets of CD8+ and CD4+ Ly-6C+ T cells, while Ly-6.1 strains (eg, A, BALB/c, CBA, C3H/He, DBA/1, NZB) have only CD8+ Ly-6C+ T cells.  Upregulation of Ly-6C expression on CD8+ T cells by interferons α and β and poly (I:C) has been described, and Ly-6C is a memory marker on CD8+ T cells. The HK1.4 antibody does not reportedly block the binding of the RB6-8C5 monoclonal antibody that specifically recognizes Mouse Ly-6G and Ly-6C.

      568954 Rev. 1
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      568954 Rev.1
      Citations & References
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      View product citations for antibody "568954" on CiteAb

      Development References (3)

      1. Havran WL, Lancki DW, Moldwin RL, Dialynas DP, Fitch FW. Characterization of an anti-Ly-6 monoclonal antibody which defines and activates cytolytic T lymphocytes.. J Immunol. 1988; 140(4):1034-42. (Immunogen: Activation, Blocking, (Co)-stimulation, Flow cytometry, Fluorescence activated cell sorting, Functional assay, Stimulation). View Reference
      2. Ma C, Kapanadze T, Gamrekelashvili J, Manns MP, Korangy F, Greten TF. Anti-Gr-1 antibody depletion fails to eliminate hepatic myeloid-derived suppressor cells in tumor-bearing mice.. J Leukoc Biol. 2012; 92(6):1199-206. (Clone-specific: Flow cytometry). View Reference
      3. Schlueter AJ, Malek TR, Hostetler CN, Smith PA, deVries P, Waldschmidt TJ. Distribution of Ly-6C on lymphocyte subsets: I. Influence of allotype on T lymphocyte expression.. J Immunol. 1997; 158(9):4211-22. (Biology). View Reference
      568954 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.