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      PE Rat Anti-Mouse Siglec-H
      PE Rat Anti-Mouse Siglec-H
      Multicolor flow cytometric analysis of Siglec-H expression on mouse leucocytes. BALB/c mouse splenocytes (Top Plots; preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody/Mouse BD Fc Block™, Cat. No. 553141/553142) and thymocytes (Bottom Plots) were stained with APC Rat anti-Mouse CD45R/B220 antibody (Cat. No. 553092/561880) and either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plots) or PE Rat Anti-Mouse Siglec-H (Cat. No. 567252; Right Plots) 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 Siglec-H (or Ig Isotype control staining) versus CD45R/B220 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™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      PE Rat Anti-Mouse Siglec-H
      Multicolor flow cytometric analysis of Siglec-H expression on mouse leucocytes. BALB/c mouse splenocytes (Top Plots; preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody/Mouse BD Fc Block™, Cat. No. 553141/553142) and thymocytes (Bottom Plots) were stained with APC Rat anti-Mouse CD45R/B220 antibody (Cat. No. 553092/561880) and either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plots) or PE Rat Anti-Mouse Siglec-H (Cat. No. 567252; Right Plots) 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 Siglec-H (or Ig Isotype control staining) versus CD45R/B220 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™ 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™
      Siglec-H; Siglech; sialic acid binding Ig-like lectin H
      Mouse (QC Testing)
      Rat IgG1, κ
      Mouse Extracellular Siglec-H Domain Recombinant Protein
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. 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.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      567252 Rev. 1
      Antibody Details
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      551

      The 551 monoclonal antibody specifically recognizes Siglec-H, a type I transmembrane glycoprotein that is encoded by Siglech (Sialic acid binding Ig-like lectin H). Siglec-H contains two immunoglobulin domains in its extracellular region and a cytoplasmic domain that lacks tyrosine-based signaling motifs unlike other CD33-related Siglec-like molecules. Siglec-H is expressed on progenitors of plasmacytoid dendritic cells (pDCs) and depends on DAP12 for cell surface expression and intracellular signaling function. In addition to its expression on plasmacytoid dendritic cells (pDC), Siglec-H may also be expressed by splenic marginal zone macrophages, medullary macrophages in lymph nodes, and microglia. Siglec-H may regulate the production of type I interferons (IFN-I) by pDCs and other cell types. There is no clear human ortholog for mouse Siglec-H.

      567252 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
      567252 Rev.1
      Citations & References
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      View product citations for antibody "567252" on CiteAb

      Development References (7)

      1. Blasius A, Vermi W, Krug A, Facchetti F, Cella M, Colonna M. A cell-surface molecule selectively expressed on murine natural interferon-producing cells that blocks secretion of interferon-alpha.. Blood. 2004; 103(11):4201-6. (Biology). View Reference
      2. Blasius AL, Giurisato E, Cella M, Schreiber RD, Shaw AS, Colonna M. Bone marrow stromal cell antigen 2 is a specific marker of type I IFN-producing cells in the naive mouse, but a promiscuous cell surface antigen following IFN stimulation.. J Immunol. 2006; 177(5):3260-5. (Immunogen). View Reference
      3. Koda Y, Nakamoto N, Chu PS, et al. Plasmacytoid dendritic cells protect against immune-mediated acute liver injury via IL-35.. J Clin Invest. 2019; 129(8):3201-3213. (Clone-specific: Flow cytometry). View Reference
      4. Schmitt H, Sell S, Koch J, et al. Siglec-H protects from virus-triggered severe systemic autoimmunity. J Exp Med. 2016; 213(8):1627-1644. (Clone-specific). View Reference
      5. Swiecki M, Wang Y, Gilfillan S, Lenschow DJ, Colonna M. Cutting edge: paradoxical roles of BST2/tetherin in promoting type I IFN response and viral infection.. J Immunol. 2012; 188(6):2488-92. (Clone-specific: Fluorescence activated cell sorting). View Reference
      6. Uematsu T, Iizasa E, Kobayashi N, Yoshida H, Hara H. Loss of CARD9-mediated innate activation attenuates severe influenza pneumonia without compromising host viral immunity.. Sci Rep. 2015; 5:17577. (Clone-specific: Flow cytometry). View Reference
      7. Zhang J, Raper A, Sugita N, et al. Characterization of Siglec-H as a novel endocytic receptor expressed on murine plasmacytoid dendritic cell precursors. Blood. 2006; 107(9):3600-3608. (Biology). View Reference
      View All (7) View Less
      567252 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.