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BV510 Rat Anti-Mouse Siglec-H
BV510 Rat Anti-Mouse Siglec-H
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 BD Horizon™ BUV395 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 563793) and with either BD Horizon™ BV510 Rat IgG1, κ Isotype Control (Cat. No. 563039; Left Plots) or BD Horizon™ BV510 Rat Anti-Mouse Siglec-H antibody (Cat. No. 567816; 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.
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 BD Horizon™ BUV395 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 563793) and with either BD Horizon™ BV510 Rat IgG1, κ Isotype Control (Cat. No. 563039; Left Plots) or BD Horizon™ BV510 Rat Anti-Mouse Siglec-H antibody (Cat. No. 567816; 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.
Product Details
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BD Horizon™
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 BSA and ≤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.

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 cells and CompBead 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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Violet 510 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567816 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.

567816 Rev. 1
Format Details
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BV510
The BD Horizon Brilliant Violet™ 510 (BV510) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye with an excitation maximum (Ex Max) at 327-nm / 405-nm and an emission maximum (Em Max) at 512-nm. BV510, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 510-nm (e.g., a 525/50 bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV510
Violet 405 nm
327 nm, 405 nm
512 nm
567816 Rev.1
Citations & References
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View product citations for antibody "567816" 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. (Biology: Flow cytometry). 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
567816 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.