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Multicolor flow cytometric analysis of Siglec-H expression on mouse splenocytes and thymocytes. BALB/c mouse splenic leucocytes (Top Plots; preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142] or thymocytes (Bottom Plots) were stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092) and with either BD Horizon™ BUV395 Rat IgG1, κ Isotype Control (Cat. No. 564059; Left Plots) or BD Horizon™ BUV395 Rat Anti-Mouse Siglec-H antibody (Cat. No. 567814; Right Plots) at 1.0 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-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.
BD Horizon™ BUV395 Rat Anti-Mouse Siglec-H
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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
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Companion Products
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.
Development References (7)
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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
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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. (Biology: Flow cytometry). View Reference
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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
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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
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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
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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
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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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.