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Flow cytometric analysis of Siglec-1 (CD169) expression on fresh or Interferon-α (IFN-α)-treated human peripheral blood mononuclear cells. Fresh human PBMC (Top Panels) or PBMC cultured with recombinant human IFN-α2a protein (20 ng/mL) for 48 hoursor (Bottom Panels) were prepared. The cells were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220), and stained with APC Mouse Anti-Human CD14 antibody (Cat. No. 555399/561708/561383) and either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; Left Panels) or BD Horizon BB515 Mouse Anti-Human Siglec-1 (CD169) antibody (Cat. No. 565353; Right Panels). Two-color flow cytometric contour plots showing the correlated expression of Siglec-1 (CD169) (or Ig Isotype control staining) versus CD14 were derived from gated events with the forward and side light-scatter characteristics of viable mononuclear cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BB515 Mouse Anti-Human Siglec-1 (CD169)
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 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 Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. 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).
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 7-239 monoclonal antibody specifically binds to Sialic acid-binding Ig-like lectin 1 (Siglec-1), which is also known as Sialoadhesin (SN), or CD169. Siglec-1 is a type I transmembrane glycoprotein that belongs to the Siglec family within the Ig superfamily. This adhesion molecule especially binds to glycolipids and glycoproteins with terminal α-2 sialyl residues. Siglec-1 is expressed by macrophages and dendritic cells and serves as a cellular interaction molecule. Its expression can be upregulated by cells in response to type II collagen, or to cytokines including interferons, and tumor necrosis factor. Siglec-1 plays roles in endocytosis, hematopoiesis, and leucocyte migration. It mediates macrophage binding to various cell types including developing and mature leucocytes. Siglec-1 that is expressed by dendritic cells can also bind HIV-1 and may mediate viral transfer to bystander CD4+ T cells. Several Siglec-1 counter-receptors have been described including CD43, CD206, and CD227 which are expressed by T cells, macrophages, or breast cancer cells, respectively. The 7-239 antibody reportedly blocks Siglec-1 functions in some cellular assay systems.
The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.
Development References (6)
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Hartnell A, Steel J, Turley H, Jones M, Jackson DG, Crocker PR. Characterization of human sialoadhesin, a sialic acid binding receptor expressed by resident and inflammatory macrophage populations. Blood. 2001; 97(1):288-296. (Biology). View Reference
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Izquierdo-Useros N, Lorizate M, Puertas MC, et al. Siglec-1 is a novel dendritic cell receptor that mediates HIV-1 trans-infection through recognition of viral membrane gangliosides. PLoS ONE. 10(12)(Biology). View Reference
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Kirchberger S, Majdic O, Steinberger P, et al. Human rhinoviruses inhibit the accessory function of dendritic cells by inducing sialoadhesin and B7-H1 expression. J Immunol. 2005; 175(2):1145-1152. (Immunogen: Blocking, Cell separation, Flow cytometry, Functional assay, Inhibition, Western blot). View Reference
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Rose T, Grützkau A, Hirseland H, et al. IFNα and its response proteins, IP-10 and SIGLEC-1, are biomarkers of disease activity in systemic lupus rythematosus. Arthritis Rheum. 2013; 72(10):1639-1645. (Biology). View Reference
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Xiong YS, Cheng Y, Lin QS, et al. Increased expression of Siglec-1 on peripheral blood monocytes and its role in mononuclear cell reactivity to autoantigen in rheumatoid arthritis. Rheumatology (Oxford). (Biology). View Reference
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van den Berg TK, Nath D, Ziltener HJ, et al. Cutting edge: CD43 functions as a T cell counterreceptor for the macrophage adhesion eceptor sialoadhesin Siglec-1) . J Immunol. 2001; 166(6):3637-3640. (Biology). View Reference
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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.